Pulse drug nebulization system, formulations therefore, and methods of use

ABSTRACT

Liquid nebulizer apparatus, systems, and formulation compositions, as well as systems for the nebulized, aerosol delivery of such compositions, for the administration and insufflation of medicinal aerosols into the pulmonary system of a mammal are described. The nebulizing apparatus and system can effectively aerosolize a variety of viscosities of medicinal liquid drug carriers, including those made up of oil, water, or emulsions of oil and water. Drugs dissolved or suspended in the compositions and formulations described and adapted for use herein are not damaged or denatured by the nebulization process when the nebulizer described is used. Further, the nebulization system itself can be adapted for use with both mechanically assisted pulmonary ventilation systems as well as hand-held inhalers and nose/mouth face masks for use in pulmonary drug delivery.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a divisional of, and claims priority to, U.S.patent application Ser. No. 11/855,870 filed Sep. 14, 2007, nowabandoned, which in turn claims priority to U.S. Provisional PatentApplication Ser. No. 60/845,087, filed Sep. 15, 2006, and U.S.Provisional Patent Application Ser. No. 60/891,128, filed Feb. 22, 2007,the contents of all of which are incorporated herein by reference intheir entirety.

STATEMENT REGARDING GOVERNMENT INTERESTS

The government may own rights in the present invention pursuant to grantnumber 1 P01 GM066312-01A2 from the National Institute of Health.

FIELD OF THE INVENTION

The present invention relates to compositions, formulations and methodsfor the treatment and prevention of cytoxicity, cell damage andinflammation in the pulmonary system of patients, as well as systems foruse in delivering such compositions and formulations to the patient.More particularly, the present invention relates to tocopherolcompositions and formulations for use in the treatment of pulmonarydisorders in patients, methods for their use, and systems for thedelivery of such tocopherol-containing compositions and formulations tothe pulmonary region of a patient.

BACKGROUND OF THE INVENTION

Burn inhalation injury of the lungs increases morbidity and mortality,with 70% of victims of smoke inhalation dying within the first 12 hoursof [Shirani, K. Z., et al., Ann. Surg., Vol. 205(1): pp. 82-87 (1987)].Lung injury is traumatic, and is typically caused by heat and chemicalirritation, with chemical injury being the leading lethal cause of smokeinhalation injury. Similarly, thermally injured patients who sustaininhalation injury have a 20 fold increase in mortality [see, Saffle, J.R., et al., J. Burn Care Rehabil., Vol. 16(3, pt. 1); pp. 219-232(1995)].

Injury from burn and smoke inhalation has been demonstrated to produce asystemic inflammatory response and increase levels of reactive oxygenspecies (ROS) [Traber, D. L., et al., Burns Inc. Therm. Inj., Vol.14(5): pp. 357-364 (1988)]. ROS produces an increase in pulmonarymicrovasculature and pulmonary edema accompanied by increased lipidperoxidation in lung tissue. Inhibition of lipid peroxidation has beendemonstrated to reduce these symptoms in animals subjected to hotsawdust smoke by Z. Min, et al., [J. Med. Cell. PLA, Vol. 5(2): pp.176-180 (1990)].

Antioxidants are compounds that reduce oxidation products and have beendemonstrated to reduce cytotoxicity in smoke inhalation-lung injury,adult respiratory distress syndrome, emphysema and asthma. More recentlyit has been reported that the use of antioxidants such as vitamin E maybe beneficial in the treatment of victims of fire accidents who sustainboth thermal injury to the skin and smoke inhalation and exhibitevidence of oxidant injury [Morita, N., et al., Shock, Vol. 25(3): pp.277-282 (2006)]. For example, vitamin C and vitamin E (alpha-tocopheroland gamma tocopherol) are antioxidants in vivo which may act together toscavenge ROS to produce non-reactive compounds within the human body.One of the important chemical features of the tocopherols is that theyare redox agents which act under certain circumstances as antioxidants.In acting as an antioxidant, tocopherols presumably prevent theformation of toxic oxidation products, such as perioxidation productsformed from unsaturated fatty acids. Further, it has recently beendiscovered that individual members in the class of tocopherols mayexhibit different biological properties from one another despite theirstructural similarity. Some investigators, for example, believe thatγ-tocopherol, unlike α-tocopherol, acts in vivo as a trap formembrane-soluble electrophilic nitrogen oxides and other electrophilicmutagens [Christen, S., et al. Proc. Natl. Acad. Sci. 94: 3217-3222(1997]. Vitamin E is remarkably safe, and falls within a class ofcompounds that are “generally regarded as safe” or “GRAS”. Vitamin E isavailable in several forms that present varied activities between them.Whereas alpha-tocopherol has been widely investigated for therapeuticuses, until recently gamma-tocopherol (a form of “des-methyltocopherol”) has received much less attention in science. However,gamma-tocopherol presents a variety of beneficial advantages overalpha-tocopherol in various considerations. In one particular regard,gamma-tocopherol has been characterized to exhibit much more potentanti-oxidant qualities, resulting in a unique anti-inflammatory activitynot shared with the alpha-tocopherol. In addition, gamma-tocopherol isbelieved to enhance outcomes of therapy when combined with certain otherbioactive agents or drugs. Antioxidants, and in particular, gammatocopherol is capable of preserving the elastase inhibitor capacity ofthe lower respiratory tract fluid of mammals exposed to harmful,chemical gases. Thus, it is believed that the direct delivery ofantioxidants such as vitamin E, and particularly, gamma tocopheroldirectly to the airways of mammals, may reduce or treat the injuryresulting from burn and smoke inhalation, as well as other pulmonarydisorders.

Inhalation-based therapies have been extensively evaluated assite-specific method to treat pulmonary disorders due to their abilityto rapidly and selectively deposit agents in the lung in greater amountsthan can be readily achieved by other methods [see: Kuhn, R. J.,Pharmacotherapy, Vol. 22: pp. 80S-85S (2002)]. Consequently, a varietyof aerosolized compounds have been researched and their aerosolizationattempted, including recombinant proteins, glutathione, and vitaminssuch as Vitamin E [Hybertson, B. M., et al., Free Radic. Biol. Med.,Vol. 18: pp. 537-542 (1995)]. However, these attempts have been largelyunsuccessful due to the substantial insolubility of these therapeuticagents and potential therapeutic agents in carrier systems that aresuitable for use in aerosol therapeutic delivery systems. For example,many of the pharmaceutical compounds, vitamins, and biological agentsthat exhibit promise in the treatment of pulmonary diseases, disordersand damage that result from smoke inhalation are insoluble in water andother the carriers for aerosol formulation and for use in nebulizers.Further, these same compounds which exhibit potential therapeuticapplicability in the pulmonary region of patients are often soluble onlyin oil-based solvents or compounds, and as such they are unable to beaerosolized by the current nebulizer systems available and in use. Inthe event that such oil-based formulations can be realized, and they canbe transformed into an aerosol by a nebulizer, the exceedingly high flowrate required to aerosolize them, and the resultant particle size of theaerosols makes the formulations unsuitable for use in treatment.Consequently, not only are new formulations necessary, but a newnebulizer design is required in order to convert formulations of suchtherapeutic agents into aerosols having the desired particle size in thedesired range of 2 μm to 12 μm.

The present invention meets these needs by providing novel,pharmaceutical compositions of tocopherols, such as gamma tocopherol,and tocopherol derivatives which are demonstrated herein to protectanimals from cytotoxic injury and death, pulmonary injury, as well asother injuries and disease conditions, including inflammatory diseases,as well as methods and systems for delivering these compositions by wayof nebulizing such water-insoluble drug formulations.

BRIEF SUMMARY OF THE INVENTION

In one aspect of the present disclosure, an apparatus for nebulizing anon-aqueous liquid is described.

In accordance with another aspect of the present disclosure, anebulization system comprising a nebulizing nozzle capable of nebulizinga composition comprising fatty acids or lipids is described, wherein thenebulizing nozzle comprises an outer gas-delivery tube and an innermicrochannel delivery tube having a central fluid channel, wherein theouter delivery tube is concentrically configured around the innerdelivery tube, such concentric configuration forming an annularintermediate space between the two tubes, and wherein the intermediateair space between the two tubes is the free air opening value, the valueof which is the internal diameter of the outer gas-delivery tube minusthe total inside diameter of the inner microchannel delivery tube.

In accordance with another aspect of the present disclosure, a pulmonarydrug delivery system capable of nebulizing a composition comprising awater-insoluble or substantially water-insoluble drug and a fatty acidor lipid is described, wherein the system comprises a reservoir forcontaining the drug composition; a reservoir for containing a propellantgas; a mechanical control valve capable of regulating the flow of thepropellant gas and the drug composition; and a nebulizing nozzle adaptedto receive both the drug composition and the propellant gas, wherein thenebulizing nozzle can produce aerosol droplets having a particle sizeranging from about 2 μm to about 12 μm in median mass aerodynamic size.

In another aspect of the present disclosure, formulations fornebulization of water-insoluble drugs are provided.

In another aspect of the present disclosure, methods of deliveringformulations of water-insoluble drugs via nebulizer are provided.

In one aspect of the present disclosure, a pharmaceutical kit foraerosol administration of a medicament is described.

In a further aspect of the present disclosure, methods for preparingformulations for nebulization are described.

In yet another aspect of the present disclosure, the use of antioxidantsin combination with one or more fatty acids or lipids in the preparationof a medicament comprising gamma tocopherol for the treatment ofpulmonary disorders is described.

In another aspect of the present disclosure, the use of compositionscomprising gamma-tocopherol are described for use in reducing the levelsof reactive oxygen species in the pulmonary microvasculature of apatient.

In a further aspect of the present disclosure, the use of alpha orgamma-tocopherol in the manufacture of a medicament for the inhibitionof lipid peroxidation in the pulmonary system of a patient is described.

In further aspects of the present disclosure, a non-aqueous medicinalaerosol composition is described, the composition comprising atherapeutically effective amount of an inhibitor of c-GMP-specificphosphodiesterase (PDE) type IV or type V, or a derivative, metabolite,solvate, prodrug, or polymorph thereof, and a lipid or fatty acid. Inaccordance with this aspect of the present disclosure, the inhibitor ofc-GMP PDE IV or PDE V may be sildenafil, or a derivative, metabolite,prodrug, polymorph, or solvate thereof, in a therapeutically-effectiveamount ranging from about 1 mg/kg/day to about 1,000 mg/kg/day.

In accordance with further aspects of the present disclosure, a drugnebulizing apparatus adapted for pulmonary inhalation and delivery ofaerosolized medicaments into a mammalian patient is described, whereinthe apparatus comprises a reservoir containing a drug formulation and anebulizing nozzle adapted to be fit to a breathing circuit and furtherattached to a face mask, wherein the nebulizing nozzle producesaerosolized droplets sized for pulmonary, inhaled drug delivery of thedrug formulation, and wherein the nebulizing nozzle comprises a fluidmicro-tube with an air delivery tube capable of nebulizing a fluid fromthe micro-tube into droplets into droplets sized for inhaled drugdelivery. In accordance with this aspect of the disclosure, the drugformulation may be in the form of a mixture, a solution, a suspension,or an emulsion, and the droplets may range in size from about 2 μm toabout 10 μm, including from about 2 μm to about 5 μm. In furtheraccordance with this aspect of the disclosure, the face mask istypically capable of being fit to the patient's nose, mouth, or both thenose and the mouth.

In accordance with yet another aspect of the present disclosure, a facemask for use in a pressurized drug delivery system is provided, the facemask comprising an at least partially deformable body having a surfacefor placement against a face of a patient and a nose bridge sectionformed in an upper section of the body, a vent to the atmosphere outsideof the face mask, and a connector integral to a portion of the mask, theconnector defining a fluid pathway into an interior portion of the facemask and constructed to receive, under pressure, an aerosolized drugcomposition. In association with this aspect, the face mask may becoupled to a nebulizer drug delivery system for delivering anaerosolized drug through the face mask. Further, the body of the facemask may include a bottommost surface for contacting the face when theface mask is applied against the face and the body at least partiallydeforms.

In one embodiment, the pharmaceutical compositions of the presentinvention comprises gamma tocopherol measured at about 5% to about 10%(w/v). In one embodiment, pharmaceutical compositions of the presentinvention comprise gamma tocopherol measured at about 10% to about 15%(w/v). In one embodiment, pharmaceutical compositions of the presentinvention comprise gamma tocopherol measured at about 15% to about 20%(w/v). In one embodiment, pharmaceutical compositions of the presentinvention comprise gamma tocopherol measured at about 20% to about 25%(w/v). In a preferred embodiment, the pharmaceutical compositions of thepresent invention comprise gamma tocopherol measured at about 10% (w/v).

In one embodiment, methods of preparing pharmaceutical compositions ofthe present invention further comprise adjusting the osmolarity of thepharmaceutical composition to an osmolarity in the range from about 200to about 400 mOsmol/L. In one embodiment, the osmolarity of thepharmaceutical composition is in the range from about 240 to about 360mOsmol/L or an isotonic range.

In one embodiment of the present disclosure, the pH of the tocopherolcompositions, in particular the gamma tocopherol composition, is in therange from about 2 to about 9, while in other embodiments, the pH may bein the range from about 3 to about 8. The pH of the pharmaceuticalcomposition may be adjusted to a physiologically compatible range. Forexample, in one embodiment, the pH of the pharmaceutical compositionsdescribed herein may be in the range from about 3.0 to about 7.5. Inanother embodiment, the pharmaceutical compositions of the presentinvention may have a pH in the range from about 3.5 to about 7.5.

In one embodiment, storage of the gamma tocopherol pharmaceuticalcomposition is about three months, and the storage temperature is in therange from about 15° C. to about 30° C., and more preferably in therange from about 20° C. to about 25° C. In another embodiment, storageof the gamma tocopherol pharmaceutical composition is about six months,and the storage temperature is in the range from about 15° C. to about30° C., and more preferably in the range from about 20° C. to about 25°C. In another embodiment, storage of the gamma tocopherol pharmaceuticalcomposition is about twelve months, and the storage temperature is inthe range from about 15° C. to about 30° C., and more preferably in therange from about 20° C. to about 25° C.

The present invention further includes kits comprising tocopherol andtocopherol compositions in accordance with the present disclosure. Inaccordance with further aspects of this embodiment, the presentdisclosure contemplates and includes kits comprising gamma tocopheroland γ-tocopherol pharmaceutical compositions of the present invention.In certain embodiments, such kits may comprise one or more containers tostore the gamma tocopherol pharmaceutical compositions.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

The following figures form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these figures in combination with the detailed description ofspecific embodiments presented herein.

FIG. 1A. illustrates a cross-sectional view of a micro-channelnebulizing nozzle in accordance with an aspect of the presentdisclosure.

FIG. 1B. illustrates a partial cut-away view of the proximal end of thenebulizing nozzle illustrated in FIG. 1A.

FIG. 1C illustrates an end-view of the proximal end of the nebulizingnozzle of FIG. 1B, taken along line 1-1.

FIG. 2A illustrates a schematic side view of a portion of a nebulizer inaccordance with an aspect of the present disclosure.

FIG. 2B illustrates a cross-sectional schematic view of the nebulizer ofFIG. 2A, taken along line 2-2.

FIG. 3 illustrates a multiple nebulizer nozzle configuration, inaccordance with an aspect of the present disclosure.

FIG. 4 illustrates a schematic view of a nebulizing system of thepresent disclosure for use in association with a mechanical ventilator.

FIG. 5 illustrates a hand-held micro-channel nebulizing system inaccordance with an aspect of the present disclosure.

FIG. 6A illustrates a micro-channel nebulizer configured to operatecontinuously using any standard air/oxygen source commonly available foruse in respiratory therapy, for use in association with aspects of thisdisclosure.

FIG. 6B illustrates an exemplary manner in which the device of FIG. 6Amay be adapted to a semi-closed passive breathing circuit.

FIG. 7A illustrates the effect of alpha- and gamma-tocopherol onpulmonary gas exchange evaluated by measuring the PaO₂/FiO₂ ratio.

FIG. 7B illustrates the effect of alpha- and gamma-tocopherol on changesin lung lymph flow over time.

FIG. 7C illustrates the effect of alpha- and gamma-tocopherol asdelivered via the nebulizers of the present disclosure on the pulmonaryvascular permeability of a shunt fraction of animals tested, asevaluated by measuring the pulmonary shunt fraction (Qs/Qt) over time.

FIG. 7D illustrates the effect of formulations administered inaccordance with the present disclosure on peak airway pressures overtime.

FIG. 8 illustrates nebulized fatty acid droplets generated using systemsand methods of the present disclosure, impacted on a counting slide.

FIG. 9 illustrates the exemplary gamma-tocopherol concentration in lungtissue.

FIG. 10A illustrates the effect of gamma-tocopherol (γ-T) nebulizationon PaO₂/FiO₂.

FIG. 10B illustrates the effect of gamma-tocopherol (γ-T) nebulizationon pulmonary shunt fractions.

FIG. 11 illustrates the effect of γ-T nebulization on lung lymph flow(pulmonary transvascular fluid flux).

FIG. 12A illustrates the effect of γ-T nebulization on lung wet-to-dryratio.

FIG. 12B illustrates the effect of γ-T nebulization on airwayobstructions.

FIGS. 13A-13B illustrate the effect of γ-tocopherol (γ-T) nebulizationon malondialdehyde (MDA) level (A) and 3-nitrotyrosine level (B) in lungtissue.

FIGS. 14A-14B illustrate the effect of γ-tocopherol (T) nebulization onpoly(ADP-ribose) polymerase activity in lung tissue.

FIG. 15A illustrates the effect of γ-tocopherol (γ-T) nebulization onIL-8.

FIG. 15B illustrates the effect of γ-tocopherol (γ-T) on mRNA in lungtissue.

While the inventions disclosed herein are susceptible to variousmodifications and alternative forms, only a few specific embodimentshave been shown by way of example in the drawings and are described indetail below. The figures and detailed descriptions of these specificembodiments are not intended to limit the breadth or scope of theinventive concepts or the appended claims in any manner. Rather, thefigures and detailed written descriptions are provided to illustrate theinventive concepts to a person of ordinary skill in the art and toenable such person to make and use the inventive concepts.

Definitions

The following definitions are provided in order to aid those skilled inthe art in understanding the detailed description of the presentinvention.

The phrase “pharmaceutical composition” refers to a formulation of acompound and a medium generally accepted in the art for the delivery ofthe biologically active compound to mammals, e.g., humans. Such a mediumincludes all pharmaceutically acceptable carriers, diluents orexcipients therefore.

The phrase “pharmaceutically acceptable carrier, diluent or excipient”as used herein includes without limitation any adjuvant, carrier,excipient, glidant, sweetening agent, diluent, preservative,dye/colorant, flavor enhancer, surfactant, wetting agent, dispersingagent, suspending agent, stabilizer, isotonic agent, solvent, oremulsifier which has been approved by the United States Food and DrugAdministration as being acceptable for use in humans or domesticanimals.

The term “therapeutically effective amount”, as used herein, the doseadministered to an animal, such as a mammal, in particular a human,should be sufficient to prevent the targeted disease or disorder, e.g.,cancer, delay its onset, slow its progression, or treat the disease ordisorder (e.g., reverse or negate the condition). One skilled in the artwill recognize that dosage will depend upon a variety of factorsincluding the strength of the particular composition employed, as wellas the age, species, condition, and body weight of the animal. The sizeof the dose will also be determined by the route, timing, and frequencyof administration as well as the existence, nature, and extent of anyadverse side-effects that might accompany the administration of aparticular composition and the desired physiological effect.

“Biological active agent”, as used herein, refers to any amino acid,peptide, protein, or antibody, natural or synthetic, which exhibits atherapeutically useful effect. Such biologically active agents mayinclude recombinant proteins, enzymes, peptoids, or PNAs, as well ascombinations of such agents.

The phrase “pharmaceutically acceptable” or“pharmacologically-acceptable” refers to compositions that do notproduce an allergic or similar unexpected reaction when administered toa human or animal in a medical or veterinary setting.

The compositions of the present invention may be prepared forpharmaceutical administration by methods and with excipients generallyknown in the art, such as described in Remington's PharmaceuticalSciences [Troy, David B., Ed.; Lippincott, Williams and Wilkins; 21stEdition, (2005)].

“Treating” or “treatment” as used herein covers the treatment of thedisease or condition of interest, e.g., tissue injury, in a mammal,preferably a human, having the disease or condition of interest, andincludes: (i) preventing the disease or condition from occurring in amammal, in particular, when such mammal is predisposed to the conditionbut has not yet been diagnosed as having it; (ii) inhibiting the diseaseor condition, i.e., arresting its development; (iii) relieving thedisease or condition, i.e., causing regression of the disease orcondition; or (iv) relieving the symptoms resulting from the disease orcondition.

As used herein, the terms “disease,” “disorder,” and “condition” may beused interchangeably or may be different in that the particular maladyor condition may not have a known causative agent (so that etiology hasnot yet been worked out) and it is therefore not yet recognized as adisease but only as an undesirable condition or syndrome, wherein a moreor less specific set of symptoms have been identified by clinicians.

The term “water-insoluble” encompasses the terms sparinglywater-soluble, slightly or very slightly water-soluble, as well aspractically or totally water-insoluble compounds [see, Remington: TheScience and Practice of Pharmacy, vol. I, 194-195 (Gennaro, ed., 1995)].As used herein, a compound is water-insoluble for the purposes of thisinvention if it requires at least 30 parts solvent (e.g., water orsaline) to dissolve one part solute (Id.). In accordance with thepresent disclosure, the term “water-insoluble” also encompasses oil- orlipid-soluble, as well as substantially oil- or lipid soluble.

As used herein, the term “tocopherol” includes all such natural andsynthetic tocopherol or Vitamin E compounds having the general structureas shown below, including all 10 isomers (five tocopherols (α-, β-, γ-,δ-, ζ₂-) and five tocotrienols (α/ζ₁-, (β/ε-, γ-, δ-, η-), as well ascombinations thereof, including but not limited to α-tocopherol (alphatocopherol)(2,5,7,8-tetramethyl-2-(4′,8′,12′-trimethyldecyl)-6-chromanole),β-tocopherol (beta-tocopherol), γ-tocopherol (gamma tocopherol),δ-tocopherol (delta tocopherol), and ζ₂-tocopherol, as well as the d, land dl [also referred to equivalently as the (+), (−), and (±) forms]enantiomers, prodrugs, esters, solvates, and/or polymorphs thereof, ormixtures of any of these compounds. These can be represented generallyby the structure (I) below,

wherein, R₁, R₂ and R₃ may each alternatively be selected from hydrogen,alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, and heteroaralkylmoieties, any of which can be unsubstituted or substituted with one ormore of the same or different substituents, which are typically selectedfrom —X, —R′, ═O, —OR′, —SR′, ═S, —NR′R′, —NR′R′R′⁺, ═NR′, —CX₃, —CN,—OCN, —SCN, —NCO, —NCS, —NO, —NO₂, ═N₂, —N₃, —S(O)₂OH⁻, —S(O)₂OH,—S(O)₂R′, —C(O)R′, —C(O)X, —C(S)R′, —C(S)X, —C(O)OR′, —C(O)O⁻, —C(S)OR′,—C(O)SR′, —C(S)SR′, —C(O)NR′R′, —C(S)NR′R′ and —C(NR)NR′R′, where each Xis independently a halogen (F, Cl, Br, or I, preferably F or Cl) andeach R′ is independently hydrogen, alkyl, alkenyl, or alkynyl; whereinthe wavy line, “

”, represents that the stereochemistry at this point may be in the formof the E- or Z-isomer; and “

” represents that the carbon-carbon bond may be a single or double(olefinic) bond.

While practical size limits for the various substituent groups will beapparent to those skilled in the art, generally preferred are the alkyl,alkenyl, alkynyl, aryl, heteroaryl, aralkyl, and heteroaralkyl moietiescontaining up to about 40 carbon atoms, more preferably up to about 20carbon atoms and most preferably up to about 10 carbon atoms.

As to any of the above groups that contain one or more substituents, itis understood, of course, that such groups do not contain anysubstitution or substitution patterns which are sterically impracticaland/or synthetically non-feasible. In addition, the compounds of thisinvention include all stereochemical isomers and mixtures thereofarising from the substitution of these compounds.

Except as otherwise specifically provided or clear from the context, theterm “compounds” of the invention should be construed as including the“pharmaceutically acceptable salts” thereof (which expression has beeneliminated in certain instances for the sake of brevity).

The term “gamma-tocopherol” or “γ-tocopherol”, as used herein, refers to2,7,8-trimethyl-(4,8,12-trimethyltridecyl)chroman-6-ol, alternately andequally acceptably referred to as d-gamma-tocopherol,RRR-gamma-tocopherol, 2R,4′R,8′R-gamma-tocopherol, gamma-TOH, gamma-Tand gamma-TH, and having the CAS registry number [54-28-4].

As used herein, the term “vitamin” refers to those compounds which areconsidered to be nutrients required for essential metabolic reactionswithin the body, and which are capable of acting both as catalysts andparticipants in chemical reactions within the body of mammals [Kutsky,R. J. Handbook of Vitamins and Hormones. Van Nostrand Reinhold, New York(1973); Kirk-Othmer, Encyclopedia of Chemical Technology, Third Edition,John Wiley and Sons, NY, Vol. 24:104 (1984)].

As used herein, the term “%” when used without qualification (as withw/v, v/v, or w/w) means % weight-in-volume for solutions of solids inliquids (w/v), % weight-in-volume for solutions of gases in liquids(w/v), % volume-in-volume for solutions of liquids in liquids (v/v) andweight-in-weight for mixtures of solids and semisolids (w/w), such asdescribed in Remington's Pharmaceutical Sciences [Troy, David B., Ed.;Lippincott, Williams and Wilkins; 21st Edition, (2005)].

The terms “patient” and “subject”, as used herein, are usedinterchangeably and refer generally to a mammal, and more particularlyto human, ape, monkey, rat, pig, dog, rabbit, cat, cow, horse, mouse,sheep and goat. In accordance with this definition, lung surfaces ormembranes described and referenced in accordance with this disclosurerefer to those of a mammal, preferably a human or an animal testsubject, such as a sheep.

The term “particle size” or “droplet size” is used in the context of thepresent disclosure to refer to the average diameter of particles, e.g.,drug, lipid vesicles, in a suspension, and is defined herein as the“Mass Median Aerodynamic Diameter” (MMAD) which is referenced from anequivalent aqueous solution with a density of 1.0 g/ml. As the fluiddensity decreases the real droplet diameter/volume increases andconversely. Lung deposition of a particle or droplet is primarilydependent on the MMAD of the individual particle or droplet.

The term “spray dry” refers to a nebulization method that allows for theevaporation of a solvent in part of the nebulized formulation thatresults in a smaller droplet after a time when a portion of the droplethas evaporated. Within the context of this disclosure spray drying is anessential part of the operation of handheld inhalers as well as theoperation of pharmaceutical manufacturing methods and column injectionin analytical chemistry application especially in gas chromatography.

The term “droplet” (or a tiny drop) is an individual particle from afine spray of liquid that was nebulized into an aerosol.

“Insufflation” as used herein refers to blowing or inhaling a medicinalpowder, solution or formulation into the lungs of a patient.

The term “hygroscopic” generally refers herein to a condition whereby anebulized droplet composition absorbs water from the humidity in thedroplet air stream and causes the droplet to expand and grow causing theMMAD to increase in size.

The term “drug” as used in conjunction with the present disclosure meansany compound which is biologically active, e.g., exhibits or is capableof exhibiting a therapeutic or prophylactic effect in vivo, or abiological effect in vitro.

DETAILED DESCRIPTION

One or more illustrative embodiments incorporating the inventiondisclosed herein are presented below. Not all features of an actualimplementation are described or shown in this application for the sakeof clarity. It is understood that in the development of an actualembodiment incorporating the present invention, numerousimplementation-specific decisions must be made to achieve thedeveloper's goals, such as compliance with system-related,business-related, government-related and other constraints, which varyby implementation and from time to time. While a developer's effortsmight be complex and time-consuming, such efforts would be,nevertheless, a routine undertaking for those of ordinary skill the arthaving benefit of this disclosure.

In general terms, Applicants have created nebulizer assemblies,nebulizer fluid nozzle assemblies, and methods of using such assembliesto deliver aerosols comprising one or more water-insoluble oroil-soluble drugs to the pulmonary region of a patient, for the purposeof delivering a therapeutically effective amount of the drug to thepulmonary region so as to treat a disease or disorder of the pulmonarysystem.

I. Nebulizer Assembly and Design

Turning now to the figures, FIG. 1A provides a nebulizer 10 inaccordance with an aspect of the present disclosure. Nebulizer fluidnozzle assembly 10 comprises an outer gas-delivery tube 12 and an innermicrotube 14, each of which are concentrically configured and positionedalong a central axis 15, and each having a free end at the proximal end11 of the assembly 10. Intermediate between the outer gas-delivery tube12 and the inner microtube 14 is an annular intermediate space, whichserves to convey the nebulizing carrier gas via gas entry port 5 throughthe nebulizer needle from the distal to the proximal end, whereupon itacts to nebulize the liquid within inner microtube 14 into an aerosolhaving an aerosolized particle size ranging from about 1 mm to about 10mm. In accordance with the present disclosure, the particle size may becontrolled by the spatial relationship of tubes 12 and 14 to each other,and the flow rate of the carrier gas. In typical operation, generallyspeaking, drug emulsion 19 enters the inner microtube 14 of nebulizingassembly 10 via port 4 at the distal end 13, is propelled down the fluidmicrochannel 18 via an appropriate gas, and is aerosolized intoparticles of the desired size at the proximal end 11 of the assembly.Appropriate gases for use in nebulization in accordance with the presentdisclosure include oxygen, oxygen mixtures, nitrogen, argon, helium, andpurified air, as well as combinations of these gases in variousproportions (e.g., 70% oxygen, 30% nitrogen). While the outer tube 12and the inner microtube 14 are illustrated to be substantiallycylindrical in shape, those of skill in the art will appreciate thatthey can also be of any appropriate shape, providing such shape providesthe same advantageous flow rates and particle sizes as the illustratedarrangement. Additionally, while the nebulizer 10 is illustrated tocomprise inner and outer tubes which are substantially blunt at theproximal end 11 of the assembly, it is equally acceptable for eitherouter tube 12, inner microtube 14, or both to have an outer lipcomprising an annular bevel (not shown), the angle of such a bevelranging from about 5° to about 88°.

FIG. 2A illustrates a cross-sectional view of the proximal end 11 ofnebulizer fluid nozzle assembly 10. As is apparent therein, the spacingbetween the outer surface of inner-microtube 14 and the interior surfaceof outer gas-delivery tube 12 has a diameter D₃, and this has a valueproportional to the outer diameter D₁ of inner-microtube 14. Theintermediate air space 16 between the two tubes 12 and 14 may generallybe described to be the free air opening value, the value of which is theinternal diameter of the outer gas-delivery tube, D₂, minus the totaloutside diameter of the inner microchannel delivery tube, D₁. Inaccordance with aspects of the present disclosure, the outer gasdelivery tube 12 has an inner diameter D₂ ranging from about 0.01 inchesto about 0.05 inches, and the inner microchannel delivery tube 14 has anouter diameter ranging from about 0.01 inches to about 0.04 inches. Thevalue of the intermediate air space between the two tubes ranges fromabout 0.000009 in² to about 0.001 in², and more preferably from about0.00000259 in² to about 0.001 in².

In accordance with further aspects of this disclosure, the nebulizingnozzle 11 preferably has an air volume to nebulized droplet volume ratioless than about 60,000:1, and more preferably an air volume to nebulizeddroplet volume ratio less than about 15,000:1.

The micro-channel nebulizer assemblies described herein are ideallysuited for delivery of aerosols of formulation compositions comprisingoil, bound water, or emulsion formulations via nebulization in single ormulti-phase water-in-oil or oil-in-water droplets formation.Accordingly, the nebulizer assemblies described herein, such as nozzleassembly 10, are capable of generating aerosol droplets having aparticle size ranging from about 2 μm to about 20 μm, preferably fromabout 2 μm to about 12 μm, and more preferably from about 5 μm to about10 μm. Such aerosol droplet particle sizes include about 3 μm, about 4μm, about 5 μm, about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10μm, about 11 μm, about 12 μm, about 13 μm, about 14 μm, about 15 μm,about 16 μm, about 17 μm, about 18 μm, about 19 μm, and about 20 μm, aswell as ranges between any two of these values, such as from about 4 μmto about 11 μm.

An alternative aspect of the present disclosure is illustrated in FIG.3, wherein the proximal end 71 of nebulizing nozzle 70 is shown. Inaccordance with this aspect, nebulizing nozzle 70 comprises a pluralityof individual micro-channel nozzles 74 a-74 f within a larger mass airchannel/annular intermediate space 73 within outer air-delivery tube 72.As shown therein, the plurality of individual micro-channel nozzles 74is preferably arranged around a central, longitudinal axis 75 extendingthrough outer tube 72. The plurality of individual micro-channel nozzles74 can range from about 2 to about 12 nozzles, including 3, 4, 5, 6, 7,8, 9, 10, 11 and 12 individual nozzles.

Nebulizing nozzles and assemblies, in accordance with the presentdisclosure, may be made from any number of appropriate materials,including but not limited to metals of any appropriate gauge, includingstainless steel, metal alloys, coated metals, and polymeric materials,both natural and synthetic, as well as co-polymers, homo-polymers, andter-polymers of such polymeric materials. Such polymeric materials mayinclude polyethylenes, polyurethanes, polyacryaltes, polystyrenes,polymethacrylates, amino-based polymers, cellulosic-based polymers,phenolic-based polymers, and combinations thereof. Coatings suitable foruse with the materials for manufacture of the nebulizers and nebulizingassemblies described herein include both natural and synthetic coatingmaterials, and are optionally included on the assemblies so as toenhance the flow rates, protect the outer surface of the materials, orboth.

Turning to FIG. 4, another aspect of the present disclosure isillustrated generally therein. In the embodiment shown in FIG. 4, anebulizer system in accordance with the present disclosure may beadapted to work in conjunction with a mechanical ventilator 124 and hasbeen configured to synchronize the cyclic nebulization of drug dropletaerosol mist generation at the nebulizing nozzle 111A and the injectionof that drug mist at the beginning of the inspiration cycle of theventilator and inject the drug aerosol mist directly into the ventilatorcircuit at the endotracheal tube or tracheal tube 132, and directly intothe pulmonary region of the patient 114 via air tube 113. Thisconfiguration reduces material loss (e.g., aerosol drug loss) in theventilator circuit tubing.

The nebulizer nozzle with a “pulse jet” cycle operation in accordancewith the present disclosure is configured using the ventilator breathcycle signal output 118 signaling the nebulizer controller 121, havingcycle timers 119 and 120. The nebulization of the drug 133 in aoil-based solution in accordance with aspects of the present disclosureand contained within aspirator 129 is commenced at the leading edge ofthe inspiration cycle and operates for a short period of time at thebeginning of inspiration. The output of the nebulizer is synchronizedwith the inspiration start trigger generated by the existing electroniccontrol outputs of the ventilator. With continued reference to FIG. 4,the ventilator electronic output signal 118 then signals cycle timers119 and 120 that control electric solenoid valves on the air supply 130and fluid supply 131 to the nebulizer nozzle 111A, respectively. Thenebulization process is substantially instantaneous and independent ofthe tidal volume inspiration flow volume. The nebulized particles arethen injected directly into the ventilator air stream 113 ahead of thebulk of the inspired tidal volume. The output of the nebulizing nozzle111A may be connected directly into the end of the ventilator tubing “Y”123 at the ET tube connector. The existing ventilator O₂ mixer 125output may be tapped via connection 126 to provide the nebulizeratomizing gas supply, thus providing nebulizer gas at the same O₂concentration as the ventilator inspired oxygen concentration (FiO₂).The nebulizer cycle time, about 1.2 seconds, for the air flow isslightly longer than the fluid flow cycle time, which is about 0.4seconds. The fluid flow may be delayed about 0.2 seconds from the startof the cycle and ends before the end of the air flow. This dual cycleclears the nozzle with air so the nozzle does not drip. Total drugvolume nebulized and delivered with each breath is a function of thefluid cycle duration fluid viscosity/fluid micro-channel size and thenumber of individual nebulizing nozzles.

The devices described and illustrated in FIG. 4 may be readilymanufactured from existing technologies and electronic controlconfigurations. The system uses existing signal generations from thevarious mechanical ventilation devices as the controller for thenebulizing device. The entire device simply adapts with the existingmechanical ventilating equipment.

In a further aspect of the present disclosure, a portable handheldnebulizing or inhaler device 50 is illustrated generally in FIG. 5,comprising a lower equalization chamber portion 43 and an upper pressurevessel portion 35, wherein the inhaler device 50 is capable ofdelivering metered doses of a medicament to a patient. In thisarrangement, the medicinal formulation 38 comprising the medicament,such as described in more detail below, is retained within a bladder 37,separated from the propellant gas 36 contained within a pressure vessel35. In accordance with this structural relationship, the medicinalcomposition within bladder 37 is not emulsified with the propellant gas36, thus allowing many different propellant gases to be used in thisconfiguration including those gasses that may be difficult or impossibleto solubilize with the drug formulation. Bladder 37 is attached to stem45 by an appropriate attachment means, including mechanical attachmentmeans such as a clip, chemical attachment means such asmedically-acceptable glues and adhesives, and combinations thereof. Thedrug formulation flow and gas flow is actuated mechanically. Ports 39,40 in the actuating valve assembly 41 allow the drug composition 38 andthe liquefied gas 36 to enter into the valve assembly 41 and on to thenebulizing nozzle 52. Valve assembly 41 acts to not only transfer thedrug composition from the bladder 37 to the nebulizing nozzle 52, butalso acts to connect pressure vessel 35 with lower equalization chamber43. The pressurized drug composition flows to the end of the micro-fluidchannel 9 and then is disrupted into aerosol mist 42 by the expandingpropellant gas. In one aspect of the present disclosure, it ispreferable to use a liquefied propellant gas, such as fluorinatedhydrocarbon, in this configuration in order to provide enough gas volumefor multiple actuations and also to adapt the gas vessel for a loweroperating pressure. In accordance with another aspect of the presentdisclosure, it is preferable to use a non-liquified gas as thepropellant gas, such as carbon dioxide, nitrogen, or nitrous oxide.Preferably, in accordance with this aspect, the gas is nitrogen gas,owing to its odorless and tasteless characteristics, and its substantialinsolubility in product formulations.

In yet another aspect of the present disclosure, the formulations anddevices described herein can provide for medicinal droplets that do notsubstantially evaporate. As such, a mist of persistent and size stabledroplets containing medicine can be generated into a large vessel. Ingeneral, droplets in the 2-5 μm are well known to be the optimum sizefor pulmonary drug delivery and are well known to stay suspended in airwith only minor movement of air currents within the vessel. Importantly,in this embodiment, an environment of medicinal droplets is produced ina space and remains suspended and persist as a stable aerosol in the airwithin the environment until inhaled. The method described would besuitable for passive inhaled drug delivery for one or more persons. Moreparticularly, the embodiment would provide for drug delivery for manypersons as in a mass casualty situation where a medicinal nebulizercould be shared with one or more persons. This embodiment would beuseful to minimize equipment and personnel for mass inhalation pulmonarydrug delivery.

Still, in yet another embodiment, the devices and methods describedherein may be useful in industrial applications where the generation ofan aerosol from a viscose fluid or emulsion is required. Theseapplications shall include pharmaceutical manufacture, oil micro-dropletlubrication and fuel injection.

II. Nebulizing Mask Adaptations

In accordance with further aspects of the present disclosure, and asillustrated in FIG. 6A and FIG. 6B, the micro-channel nebulizer systemsdescribed above may be adapted for continuous or semi-continuousnebulization and delivery of inspirable droplets via a semi-closed facemask breathing circuit 62. The drug is contained in a flexible membranewithin a pressurized housing, which allows for the nebulizer to operatein various positions unaffected by gravity. Humidified air is providedby a separate standard mask humidifier which is existing equipment forface mask delivery of oxygen. The mask and lipid nebulizer is intendedto be adaptable to existing equipment.

There are certain instances wherein it is desirable to provide medicinesby nebulization and subsequent inhalation by a spontaneously breathingperson. In this application, the face mask may be coupled to a nebulizerdrug delivery system for delivering an aerosolized drug through the facemask, such that medicines are continuously nebulized into a space thatholds the medicinal droplets suspended in a contained inspirable airflow which is in proximity to the mouth and nose of the person. Thenebulizer continuously generates droplets into the air volume in a tubeconnected to a close fitting mask coving the mouth and nose of theperson. As the person inhales the medicine/air mixture, the medicineenters the lungs via the mouth and nose. On exhale, a valve on the maskopens and allows the exhaled air to flow out of the system.

FIG. 6A generally illustrates a micro-channel nebulizer configured tooperate continuously using any standard (e.g., 50 psi) air/oxygen sourcecommonly available for use in respiratory therapy, for use inassociation with this aspect of the disclosure. A micro-channelnebulizing nozzle 1A may be pressed into a plastic block containing airchannels and a pressurized medicament chamber 49. Air/oxygen is suppliedthrough a hose 55 connected to the ports of the air channel 51. Air isallowed to flow through the channel 51 and on to the micro-channel airdelivery tube 8. A portion of the supplied air is regulated by a springloaded flow valve 46 and allowed to flow into and pressurize themedicine reservoir chamber 49 containing a medicine reservoir assembly56. Another spring loaded blow-off valve 47 regulates the pressure inmedicament reservoir 49. Excess pressure, as regulated by blow-off valve47, is vented through a port 48. The pressure in the reservoir exertspressure through ports 53 on the medicament bladder 37 and forces themedicine 38 in the bladder 37 into the micro-channel liquid deliverytube 9.

FIG. 6B illustrates an exemplary manner in which the continuousnebulizing device in FIG. 6A may be adapted to a semi-closed passivebreathing circuit. As illustrated therein, a face mask 62 is included,wherein the face mask is contoured to cover the nose and mouth of apatient. Humidified air 61 and nebulized medicine is combined in a “Y”adapter 60. The humidified air/medicinal mist is then fed into the facemask through port 63. The person inhales the medicine/air mixture duringthe course of normal respiration, and then exhales. The person's exhaledbreath may then be vented through one or more one-way valves 64 on mask62, allowing the exhaled breath to exit the breathing circuit withoutinhibiting the flow of nebulized medicine.

III. Formulation of Water-Insoluble Drugs

Compositions comprise water-insoluble or substantially water-insolubledrugs or biologically active substances, as well as oil-soluble orlipid-soluble drugs and biologically active substances, in combinationwith one or more lipids or fatty acids, including naturally-occurringfats and oils. In accordance with one aspect of the present disclosure,the compositions can comprise a water-insoluble or substantiallywater-insoluble drug or biologically active substance and one or morefatty acids. In a further aspect, the compositions can comprise awater-insoluble or substantially water-insoluble drug or biologicallyactive substance, one or more fatty acids, and one or more surfactants.In yet another aspect, the compositions suitable for use with thenebulizer assemblies of the present disclosure can comprise awater-insoluble or substantially water-insoluble drug or biologicallyactive substance, water, and at least one gelling agent.

The compositions, according to the invention, may be comprised of a drugitself or any mixture of a biologically active substance with a solvent,and oil, a gelling agent, a carrier or adjuvant, emulsifier, one or moredifferent drugs, polymers, excipients, coatings and combinationsthereof. In essence, the drug(s) or substances can be combined with anycombination of pharmaceutically acceptable components to be delivered tothe cellular surfaces within the pulmonary system by the methoddescribed herein, e.g., pulmonary drug delivery. The drug(s) does nothave to be dissolved in a drug delivery medium solvent but can besuspended or emulsified in a solvent or medium. The delivery medium cantake the form of an aqueous mixture, oil, or an organic liquid. Thedelivery media solution can also comprise microspheres or nanospheres ofbiologically active substances.

The compounds useful in the formulations and therapeutically-usefulcompositions of the present invention can be used in the form ofpharmaceutically acceptable salts derived from inorganic or organicacids. The term “pharmaceutically acceptable salt” as used herein ismeant to refer to those salts which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humansand lower animals without undue toxicity, irritation, allergic responseand the like and are commensurate with a reasonable benefit/risk ratio.Pharmaceutically acceptable salts are well-known in the art. Forexample, P. H. Stahl, et al. describe pharmaceutically acceptable saltsin detail in “Handbook of Pharmaceutical Salts: Properties, Selection,and Use” (Wiley VCH, Zunch, Switzerland: 2002). The salts can beprepared in situ during the final isolation and purification of thecompounds of the present invention or separately by reacting a free basefunction with a suitable organic acid. Representative acid additionsalts include, but are not limited to acetate, adipate, alginate,citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate,camphorate, camphorsulfonate, digluconate, glycerophosphate,hemisulfate, heptanoate, hexanoate, flimarate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate),lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate,oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate,pivalate, propionate, succinate, tartrate, thiocyanate, phosphate,glutamate, bicarbonate, p-toluenesulfonate and undecanoate. Also, thebasic nitrogen-containing groups can be quaternized with such agents aslower alkyl halides such as methyl, ethyl, propyl, and butyl chlorides,bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyland diamyl sulfates; long chain halides such as decyl, lauryl, myristyland stearyl chlorides, bromides and iodides; arylalkyl halides likebenzyl and phenethyl bromides and others. Water or oil-soluble ordispersible products are thereby obtained. Examples of acids which canbe employed to form pharmaceutically acceptable acid addition saltsinclude such inorganic acids as hydrochloric acid, hydrobromic acid,sulphuric acid and phosphoric acid and such organic acids as oxalicacid, maleic acid, succinic acid and citric acid.

Basic addition salts can be prepared in situ during the finalpreparation, formulation, or purification of the therapeutically-usefulcompounds, substantially water-insoluble compounds described for use inaspects of this disclosure by reacting a carboxylic acid-containingmoiety with a suitable base such as the hydroxide, carbonate orbicarbonate of a pharmaceutically acceptable metal cation or withammonia or an organic primary, secondary or tertiary amine.Pharmaceutically acceptable salts include, but are not limited to,cations based on alkali metals or alkaline earth metals such as lithium,sodium, potassium, calcium, magnesium and aluminum salts and the likeand nontoxic quaternary ammonia and amine cations including ammonium,tetramethylammonium, tetraethylammonium, methylamine, dimethylamine,trimethylamine, triethylamine, diethylamine, ethylamine and the like.Other representative organic amines useful for the formation of baseaddition salts include ethylenediamine, ethanolamine, diethanolamine,piperidine, piperazine and the like.

Pharmaceutically acceptable salts of compounds which may be used informulations and systems described herein may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (for example, sodium,potassium or lithium) or alkaline earth metal (for example calcium ormagnesium) salts of carboxylic acids can also be made.

The formulations described for use herein may conveniently be presentedin unit dosage form and may be prepared by any of the methods well knownin the art of pharmacy. All methods include the step of bringing intoassociation one or more of the compounds described herein, or apharmaceutically acceptable salt or solvate thereof (“activeingredient”), with a carrier which constitutes one or more accessoryingredients. In general, the formulations are prepared by uniformly andintimately bringing into association the active ingredient with liquidcarriers or finely divided solid carriers or both and then, ifnecessary, shaping the product into the desired formulation.

The compound or a pharmaceutically acceptable ester, salt, solvate orprodrug thereof, such as a pharmaceutically acceptable ester, salt,solvate, or prodrug of alpha-, beta-, or gamma-tocopherol, can be mixedwith other active materials that do not impair the desired action, orwith materials that supplement the desired action, including other drugsagainst inflammatory disease or lung injury.

As recited above, the present invention uses (1) a novel nebulizingnozzle configuration that requires limited energy from a compressed gasto cause droplet nebulization from a mass fluid into droplets of asuitable size; and (2) a drug carrier(s) that is based on essentialfatty acid oils, lipids, gelled aqueous solutions, emulsions, orcombinations thereof that are harmlessly absorbed by the lung tissuesand are metabolized or expired.

Water-insoluble, substantially water-insoluble drugs, or sparingly watersoluble drugs suitable for use in the medicament compositions of thepresent disclosure include, but are not limited to, vitamins,antioxidants, anti-bronchitus agents, anti-pneumonia agents, pulmonaryanti-cancer agents, antianginal agents, antihypertensive agents, andcombinations thereof. In accordance with one aspect of the presentdisclosure, the preferred drug is a vitamin or a combination of two ormore vitamins. Suitable vitamins for use herein include but are notlimited to Vitamin A, Vitamin B (including Vitamin B₁₂), Vitamin C,Vitamin D, Vitamin E, Vitamin K3 (menadione;1,4-dehydro-1,4-dioxo-2-methyl-naphthalene, MNQ), retinol, riboflavin,niacin, ascorbic acid, β-carotene, and Coenzyme Q, including variousderivatives of Coenzyme Q having various isoprenoid side chains,including but not limited to QH, QH₂, Q₃ and Q₁₀.

In accordance with a further aspect of the present disclosure, theformulation composition suitable for nebulization using the assembliesdescribed herein comprises Vitamin E, also known as tocopherol, or aderivative, metabolite, ester, hydrate, solvate, prodrug, or polymorphthereof. Tocopherols suitable for use in the compositions herein includeall those tocopherols within the range of natural and syntheticcompounds known by the generic term Vitamin E. In accordance with oneaspect of the present disclosure, the tocopherol may be selected fromthe group consisting of alpha-tocopherols, beta-tocopherols,gamma-tocopherols, and delta tocopherols, as well as combinationsthereof. Tocopherols occur in a number of isomeric forms, the D and DLforms being most widely available, all of which are suitable for useherein. In accordance with one aspect of the present disclosure, thetocopherols are preferably gamma-tocopherols.

Tocopherols suitable for use in accordance with these aspects of thepresent disclosure may be obtained from plants, such as by extractedfrom plants using known procedures, or prepared synthetically usingknown organic synthetic methods, all of which are in accordance with thepresent disclosure. Additionally, and as suggested above, any of theforms or isomers of tocopherols and their derivatives, eg. esters may beused according to the present invention. Thus for example,gamma-tocopherol can be used as such, or in the form of its esters suchas gamma-tocopherol acetate, linoleate, nicotinate or hemisuccinate-ester, many of which are available commercially or throughknown synthetic routes.

In accordance with further aspects and embodiments of the presentdisclosure, the compositions may comprise antianginal and/orantihypertensive drugs or biologically active agents which are insolubleor substantially insoluble in water, or exhibit poor water solubility(e.g., less than about 5 mg/mL). Compounds of these types suitable foruse herein include inhibitors of cAMP (3′,5′-cyclic adenosinemonophosphate), cGMP (3′,5′-cyclic guanosine monophosphate), inhibitorsof cGMP-specific phosphodiesterase type IV (PDE IV), inhibitors ofc-GMP-specific phosphodiesterase type V (PDE V), drugs that may exhibitanti-anginal effects, including drugs which exhibit therapeutic effectson angina pectoris, and compounds which can enhance the natriureticeffect of atrial natriuretic peptide (ANP). Suitable examples of suchdrugs include but are not limited to atenolol, amlodipine, diltiazem,eplerenone, naphthyriclin-4-one derivatives, griseolic acid,dihydrodesoxygriseolic acid, derivatives of griseolic acid anddihydrodesoxygriseolic acid, angiotensin converting enzyme inhibitors,todalafil, vardenafil, ranolazine [see, Tafreshi, M. J., et al., Ann.Pharmacother., Vol. 40(4): pp. 689-693 (2006)], sildenafil, sildenafilcitrate (Viagra®; Pfizer, Inc., New York), N-desmethyl sildenafil, andT-1032(methyl-2-(4-aminophenyl)-1,2-dihydro-1-oxo-7-(2-pyridinylmethoxy)-4-(3,4,5-trimethoxyphenyl)-3-isoquionolinecarboxylate sulfate; see: Noto, T., et al., J. Pharm. Exp. Ther., Vol.294(3): pp. 870-875 (2000)], as well as derivatives, solvates, prodrugs,and polymorphs thereof. In accordance with one aspect of thisembodiment, the drug is sildenafil. Such drugs may be used in atherapeutically effective amount ranging from about 1 mg/kg/d to about1,000 mg/kg/d, as well as therapeutically effective amounts within thisrange.

“Naturally-occurring fats and oils” as used herein refers to theglyceryl esters of fatty acids (i.e., triglycerides) normally found inanimal or plant tissues, including those which have been hydrogenated toreduce or eliminate unsaturation. Naturally occurring fats and oilsinclude vegetable oils such as linseed oil, soybean oil, sunflower seedoil, corn oil, sesame oil, olive oil, castor oil, coconut oil, palm oil,peanut oil, jojoba oil, neem oil, and macadamia nut oil.

Selected naturally-occurring fats and oils suitable for use informulations of the present disclosure include, but are not limited to,the following compounds: Adansonla Digitata Oil; Apricot (Prunusarmeniaca) Kernel Oil; Argania Spinosa Oil; Argemone Mexicana Oil;Avocado (Persea gratissima) Oil; Babassu (Orbignya olelfera) Oil; BalmMint (Melissa officinalis) Seed Oil; Bitter Almond (Prunus amygdalusamara) Oil; Bitter Chemy (Prunus cerasus) Oil; Black Currant (Ribesnigrum) Oil; Borage (Borago officinalis) Seed Oil; Brazil (Bertholletiaexcelsa) Nut Oil; Burdock (Arctium lappa) Seed Oil; Butter; C12-18 AcidTriglyceride; Calophyllum Tacamahaca Oil; Camellia Kissi Oil; CamelliaOleifera Seed Oil; Canola Oil; Caprylic/Capric/Liuric Triglyceride;Caprylic/Capric/Linoleic Triglyceride; Caprylic/Capric/Myristic/StearicTriglyceride; Caprylic/Capric/Stearic Triglyceride; Caprylic/CapricTriglyceride; Caraway (Carum carvi) Seed Oil; Carrot (Daucus CarotaSativa) Oil; Cashew (Anacardium occidentale) Nut Oil; Castor OilBenzoate; Castor (Ricinus communis) Oil; Cephalins; Chaulmoogra(Taraktogenos kurzii) Oil, Chia (Salvia hispanica) Oil; Cocoa (Theobramacocao) Butter; Coconut (Cocos nucifera) Oil; Cod Liver Oil; Coffee(Coffea arabica) Oil; Corn (Zea mays) Germ Oil; Corn (Zea mays) Oil;Cottonseed (Gossypium) Oil; C10-18 Triglycerides; Cucumber (Cucumissativus) Oil; Dog Rose (Rosa canina) Hips Oil; Egg Oil; Emu Oil;Epoxidized Soybean Oil; Evening Primrose (Oenothera biennis) Oil; FishLiver Oil; Gevuina Avellana Oil; Glyceryl Triacetyl Hydroxystearate;Glyceryl Triacetyl Ricinoleate; Glycolipids; Glycosphingolipids; GoatButter; Grape (Vitis vinifera) Seed Oil; Hazel (Croylus americana) NutOil; Hazel (Corylus aveilana) Nut Oil; Human Placental Lipids; HybridSafflower (Carthamus tinctorius) Oil; Hybrid Sunflower (Helianthusannuus) Seed Oil; Hydrogenated Canola Oil; Hydrogenated Castor Oil;Hydrogenated Castor Oil Laurate; Hydrogenated Castor Oil Triisostearate;Hydrogenated Coconut Oil; Hydrogenated Cottonseed Oil; HydrogenatedC12-18 Triglycerides; Hydrogenated Fish Oil; Hydrogenated Lard;Hydrogenated Menhaden Oil; Hydrogenated Milk Lipids; Hydrogenated MinkOil; Hydrogenated Olive Oil; Hydrogenated Orange Roughy Oil;Hydrogenated Palm Kernel Oil; Hydrogenated Palm Oil; Hydrogenated PeanutOil; Hydrogenated Rapeseed Oil; Hydrogenated Shark Liver Oil;Hydrogenated Soybean Oil; Hydrogenated Tallow; Hydrogenated VegetableOil; Isatis Tinctoria Oil; Job's Tears (Coix Lacryma-Jobi) Oil; JojobaOil; Kiwi (Actinidia chinensis) Seed Oil; Kukui (Aleurites Moluccana)Nut Oil; Lard; Lauric/Palmitic/Oleic Triglyceride; Linseed (Linumusitatissiumum) Oil; Lupin (Lupinus albus) Oil; Macadamia Nut Oil;Macadamia Ternifolia Seed Oil; Macadamia Integrifolia Seed Oil; MaleatedSoybean Oil; Mango (Mangifera indica) Seed Oil; Marmot Oil; Meadowfoam(Limnanthes fragra alba) Seed Oil; Menhaden Oil; Milk Lipids; Mink Oil;Moringa Pterygosperma Oil; Mortierella Oil; Musk Rose (Rosa moschata)Seed Oil; Neatsfoot Oil; Neem (Melia azadirachta) Seed Oil; Oat (Avenasativa) Kernel Oil; Oleic/Linoleic Triglyceride;Oleic/Palmitic/Lauric/Myristic/Linoleic Triglyceride; Oleostearine;Olive (Olea europaea) Husk Oil; Olive (Olea europaea) Oil; OmentalLipdis; Orange Roughy Oil; Ostrich Oil; Oxidized Corn Oil; Palm (Elaeisguineensis) Kernel Oil; Palm (Elaeis guineensis) Oil; Passionflower(Passiflora edulis) Oil; Peach (Prunus persica) Kernel Oil; Peanut(Arachis hypogaea) Oil; Pecan (Caiya illinoensis) Oil; Pengawar Djambi(Cibotium barometz) Oil; Phospholipids; Pistachio (Pistacia vera) NutOil; Placental Lipids; Poppy (Papaver orientale) Oil; Pumpkin (Cucurbitapepo) Seed Oil; Quinoa (Chenopodium Quinoa) Oil; Rapeseed (Brassicacampestris) Oil; Rice (Oryza sativa) Bran Oil; Rice (Oryza sativa) GermOil; Safflower (Carthamus tinctorius) Oil; Salmon Oil; Sandalwood(Santalum album) Seed Oil; Seabuchthorn (Hippophae rhamnoides) Oil;Sesame (Sesamum indicum) Oil; Shark Liver Oil; Shea Butter(Butyrospermum parkii); Silk Worm Lipids; Skin Lipids; Soybean (Glycinesoja) Oil; Soybean Lipid; Sphingolipids; Sunflower (Helianthus annuus)Seed Oil; Sweet Almond (Prunus amygdalus dulcis) Oil; Sweet Chemy(Prunus avium) Pit Oil; Tali Oil; Tallow; Tea Tree (Melaleucaalternifolia) Oil; Telphairia Pedata Oil; Tomato (Solanum lycopersicum)Oil; Triarachidin; Tiibehenin; Tricaprin; Tricaprylin; TrichodesmaZeylanicum Oil; Trierucin; Triheptanoin; Triheptylundecanoin;Trihydroxymethoxystearin; Trihydroxystearin; Triisononanoin;Triisopalmitin; Triisostearin; Trilaurin; Trilinolein; Trilinolenin;Trimyristin; Trioctanoin; Triolein; Tripalmitin; Tripalmitolein;Triricinolein; Trisebacin; Tristearin; Triundecanoin; Tuna Oil;Vegetable Oil; Walnut (Juglans regia) Oil; Wheat Bran Lipids; Wheat(Triticum vulgare) Germ Oil, and combinations of such fatty acid oils.In accordance with one preferred aspect of this embodiment, theformulation composition comprises Linseed Oil, which is also known asflaxseed oil, as well as fatty acids found therein, including but notlimited to linolenic acid (LA), linoleic acid, oleic acid, stearic acid,palmitic acid, alpha-linolenic acid (LNA), and gamma-linolenic acid(GLA), any of which may be saturated or unsaturated as appropriate. Inaccordance with further aspects of the present disclosure, theformulations may also comprise hylauronic acid in an amount suitable tominimize water from transpiring across the droplets formed by thenebulizer.

Tocopherol, and in particular gamma tocopherol, compositions of thepresent invention may further optionally comprise preservatives. As usedherein, the term “preservative” is intended to mean a compound used toprevent the growth of microorganisms. Such preservatives may be used inthe tocopherol and gamma tocopherol pharmaceutical compositionsdescribed herein at typical concentrations in accordance with currentpharmaceutical practices as described. [see: The United StatesPharmacopeia—National Formulary, 29th Edition, (2006) Rockville, Md.;and, Remington's Pharmaceutical Sciences, 21st Edition, Troy, D B, Ed.Lippincott, Williams and Wilkins; (2005)]. Exemplary preservatives whichmay be used with the compositions and systems of the present disclosureinclude but are not limited to antifungal and antimicrobialpreservatives, such as benzoic acid, hydroxy benzoate and itsderivatives, butylparaben, ethylparaben, methylparaben, propylparaben,sodium benzoate, benzalkonium chloride, benzethonium chloride, benzylalcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethylalcohol, phenylmercuric nitrate and thimerosal; and antioxidants, suchas ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole,butylated hydroxytoluene, hypophosphorus acid, monothioglycerol, propylgallate, sodium ascorbate, sodium bisulfite, sodium formaldehydesulfoxylate, and sodium metabisulfite, as well as combinations of two ormore of the these preservatives.

Tocopherol and gamma tocopherol pharmaceutical compositions andformulations of the present invention may further comprise one or morepH modifying agents (buffering agents), in order to maintain the pH ofthe composition in the desired range, e.g., from a pH of from about 3.5to about 8. pH modifying agents suitable for use herein, include, butare not limited to, inorganic salts, alkali earth and/or alkali rareearth hydroxides (e.g., NaOH, KOH, or CsOH); carbonate or bicarbonate ofany appropriate alkali or alkali rare earth metal (e.g., Na₂CO₃, K₂CO₃,NaHCO₃, and KHCO₃); phosphates, such as calcium hydrogen phosphate,potassium metaphosphate, and potassium phosphate monobasic; inorganicacids such as hydrochloric acid (HCl), and organic acids such as aceticacid, citric acid, succinic acid, fumaric acid, malic acid, maleic acid,glutaric acid or lactic acid, as well as combinations thereof, any ofwhich may be water-soluble or water-insoluble, anhydrous or hydrated(e.g., dehydrate or semihydrate), as appropriate.

Others components which may be included in the therapeutically usefulcompositions of the present disclosure include but are not limited tobinding materials (e.g., block polymers, natural and synthetic rubber,polyacrylates, polyurethanes, silicones and styrene-butadienecopolymers); colorants, including but not limited to FD & C yellow #6,FD & C red #40, FD & C blue #2, and FD & C violet #1, as well as anyother appropriate dye or combination of dyes; and, UV inhibitors, toinhibit UV decomposition or isomerization of the therapeuticcompositions.

Other pharmaceutically acceptable formulation excipients may also beused in accordance with the formulation compositions described anddisclosed herein, including but not limited to coatings, stabilizers,emulsifiers, and the like, such as those described in “The Handbook ofPharmaceutical Manufacturing Formulations” [Niazi, S. K., CRC Press(2004)]. Additionally, and in accordance with aspects of the presentdisclosure, one or more surface active agents (surfactants) may be addedto the formulation compositions as appropriate. Although not required,incorporation of a compatible surfactant can improve the stability ofthe instant respiratory dispersions, increase pulmonary deposition andfacilitate the preparation of the suspension. Moreover, by altering thecomponents, the density of the particle or structural matrix may beadjusted to approximate the density of the surrounding medium andfurther stabilize the dispersion. Any suitable surface active agent(surfactant) may be used in the context of the present invention,provided that the surfactant is preferably physiologically acceptable.Physiologically acceptable surfactants are generally known in the artand include various detergents and phospholipids, as discussed in moredetail below. In accordance with one aspect, it is preferred that thesurfactant is a phospholipid including, but not limited to, an extractof a natural surfactant such as any number of known pulmonarysurfactants, including bovine- and calf-lung surfactant extracts, an eggphospholipid, a vegetable oil phospholipid such as a soybeanphospholipid, or phosphatidylcholine. Preferably, in accordance withaspects of the present disclosure, the surfactant suitable for use withthe therapeutic compositions of the present disclosure is an extract ofa natural surfactant, an egg phospholipid, or combinations thereof. Morepreferably, the compositions may any one or more of a number ofbiocompatible materials as surfactants, such as surfactants comprisingphospholipids.

In a broad sense, surfactants suitable for use in the present inventioninclude any compound or composition that aids in the formation andmaintenance of the stabilized respiratory/pulmonary dispersions byforming a layer at the interface between the particles of therapeuticcompound (e.g., tocopherol or γ-tocopherol) and the suspension medium.The surfactant may comprise a single compound or any combination ofcompounds, such as in the case of co-surfactants. In accordance withcertain aspects of the present disclosure, depending upon the specifictherapeutic composition or formulation, preferred surfactants includebut are not limited to those surfactants that are substantiallyinsoluble in the medium, nonfluorinated, and selected from the groupconsisting of saturated and unsaturated lipids, especially those thatare obtained or extracted from natural sources, nonionic detergents,nonionic block copolymers, ionic surfactants, and combinations of suchagents. It should be emphasized that, in addition to the aforementionedsurfactants, suitable (i.e. biocompatible) fluorinated surfactants arecompatible with the teachings herein and may be used to provide thedesired stabilized therapeutic preparations.

Lipids, including phospholipids, from both natural and synthetic sourcesare particularly compatible with the present inventions and may be usedin varying concentrations to form the particle or structural matrixuseful in the final, therapeutic compositions. Generally compatiblelipids include those that have a gel to liquid crystal phase transitiongreater than about 40° C. Preferably, and as described in more detailbelow, the incorporated lipids are relatively long chain (i.e. C₁₆-C₂₂)saturated lipids and preferably comprise one or more phospholipids.Exemplary phospholipids useful in the disclosed stabilized preparationsof the present invention comprise egg phosphatidylcholine,dilauroylphosphatidylcholine, dioleylphosphatidylcholine,dipalmitoylphosphatidyl-choline, disteroylphosphatidylcholine,short-chain phosphatidylcholines, phosphatidylethanolamine,dioleylphosphatidylethanolamine, phosphatidylserine,phosphatidylglycerol, phosphatidylinositol, glycolipids, gangliosideGM1, sphingomyelin, phosphatidic acid, cardiolipin; lipids bearingpolymer chains such as polyethylene glycol, chitin, hyaluronic acid, orpolyvinylpyrrolidone; lipids bearing sulfonated mono-, di-, andpolysaccharides; fatty acids such as palmitic acid, stearic acid, andoleic acid; cholesterol, cholesterol esters, and cholesterolhemisuccinate. Due to their excellent biocompatibility characteristics,phospholipids and combinations of phospholipids and poloxamers areparticularly suitable for use in the stabilized dispersions disclosedherein. With regard to biologically-compatible surfactants comprisingphospholipids, in accordance with certain aspects of the presentdisclosure, it is preferable that such biologically-compatiblesurfactants for use with the therapeutic compositions described herein,especially those comprising tocopherol and/or gamma-tocopherol, includethose surfactants that are extracts of natural surfactants, inparticular pulmonary surfactants, and synthetic synthesized mixtures ofpulmonary surfactants in order to mimic natural lung surfactant.Exemplary surfactants suitable for use with the present compositions oftocopherol and gamma-tocopherol include but are not limited to SURVANTA®(beractant, available from the Ross Products Division of AbbottLaboratories), a bovine-lung pulmonary surfactant comprisingphospholipids, neutral lipids, fatty acids, and surfactant-associatedproteins; SURFAXIN® (lucinactant, available from Discovery Laboratories,Inc.); INFASURF® (calfactant, available from Forest Pharmaceuticals,Inc., St. Louis, Mo.), an extract of natural surfactant from calf lungwhich includes phospholipids, neutral lipids, and hydrophobicsurfactant-asasociated proteins B and C(SP-B and SP-C); CUROSURF®(poractant alpha, available from Chiesi Farmaceutici, S.p.A., Parma,Italy), a non-pyrogenic pulmonary surfactant that is an extract ofnatural porcine lung comprising polar lipids (mainly phospholipids) andhydrophobic low molecular weight proteins (surfactant associatedproteins SP-B and SP-C); and ALVEOFACT® (available from BoehringerIngelheim Pharma, Ingelheim, Germany), a natural bovineextract/surfactant comprising bovine lung phospholipids; as well as thesynthetic pulmonary surfactants EXOSURF®, VENTICUTE®, ADSURF®(Pumactant™), and KL-4, all of which synthetic surfactants comprise thephospholipid dipalmitoylphosphatidylcholine (DPPC).

Compatible nonionic detergents for use in the formulations of thepresent disclosure comprise, without limitation, sorbitan estersincluding sorbitan trioleate (SPAN™ 85), sorbitan sesquioleate, sorbitanmonooleate, sorbitan monolaurate, polyoxyethylene (20) sorbitanmonolaurate, and polyoxyethylene (20) sorbitan monooleate, oleylpolyoxyethylene (2) ether, stearyl polyoxyethylene (2) ether, laurylpolyoxyethylene (4) ether, glycerol esters, and sucrose esters. Othersuitable nonionic detergents include any of those which can be easilyidentified using “McCutcheon's Emulsifiers and Detergents” (McPublishingCo., Glen Rock, N.J.) which is incorporated herein in its entirety.Preferred block copolymers include but are not limited to diblock andtriblock copolymers of polyoxyethylene and polyoxypropylene, includingpoloxamer 188 (PLURONIC™ F-68), poloxamer 407 (PLURONIC™ F-127), andpoloxamer 338. Ionic surfactants such as sodium sulfosuccinate, andfatty acid soaps may also be utilized. In accordance with certainaspects and embodiments of the present disclosure, the therapeuticcompositions described herein may comprise oleic acid or its alkalisalt.

Those skilled in the art will further appreciate that, a wide range ofsurfactants, including those not listed above, may optionally be used inconjunction with the present invention. Moreover, the optimumsurfactant, or combination thereof, for a given application can readilybe determined by empirical studies that do not require undueexperimentation. It will further be appreciated that, the preferredinsolubility of any incorporated surfactant in the suspension mediumwill dramatically decrease the associated surface activity. As such, itis arguable as to whether these materials have surfactant-like characterprior to contracting an aqueous bioactive surface (e.g. the aqueoushypophase in the lung).

On a weight to weight basis, the instant formulations and compositionsof the therapeutic compositions comprising tocopherols such asgamma-tocopherol may comprise varying levels of surfactant. In thisregard, the compositions and formulations described herein which includeone or more surfactants will preferably comprise greater than about0.1%, about 1%, about 5%, about 10%, about 15%, about 18%, or even about20% w/w % surfactant. In accordance with a further aspect of the presentdisclosure, the therapeutic compositions and formulations describedherein may comprise greater than about 25%, about 30%, about 35%, about40%, about 45%, or about 50% w/w surfactant. Still other exemplaryembodiments of the present disclosure will include therapeuticcompositions and formulations as described herein, further comprisingone or more surfactants, wherein the surfactant or surfactants arepresent at greater than about 55%, about 60%, about 65%, about 70%,about 75%, about 80%, about 85%, about 90% or even about 95% w/w.

In certain embodiments, the present invention employs a novelcomposition comprising one or more lipids associated with at least onedrug. A lipid as referred to herein is a substance that ischaracteristically insoluble in water and extractable with an organicsolvent. Lipids include, for example, the substances comprising thefatty droplets that naturally occur in the cytoplasm as well as theclass of compounds which are well known to those of skill in the artwhich contain long-chain aliphatic hydrocarbons and their derivatives,such as fatty acids, alcohols, amines, amino alcohols, and aldehydes. Ofcourse, compounds other than those specifically described herein thatare understood by one of skill in the art as lipids are also encompassedby the compositions and methods of the present invention.

A lipid for use with the present disclosure may be naturally occurringor synthetic (i.e., designed or produced by man). However, a lipid istypically a biological substance. Biological lipids are well known inthe art, and include for example and without limitation, neutral fats,phospholipids, phosphoglycerides, steroids, terpenes, lysolipids,glycosphingolipids, glycolipids, sulphatides, lipids with ether andester-linked fatty acids and polymerizable lipids, and combinationsthereof.

A. Lipid Types

A neutral fat may comprise a glycerol and a fatty acid. A typicalglycerol is a three carbon alcohol. A fatty acid generally is a moleculecomprising a carbon chain with an acidic moeity (e.g., carboxylic acid)at an end of the chain. The carbon chain may of a fatty acid may be ofany length, however, it is preferred that the length of the carbon chainbe of from about 2, about 3, about 4, about 5, about 6, about 7, about8, about 9, about 10, about 11, about 12, about 13, about 14, about 15,about 16, about 17, about 18, about 19, about 20, about 21, about 22,about 23, about 24, about 25, about 26, about 27, about 28, about 29, toabout 30 or more carbon atoms, and any range derivable therein. However,a preferred range is from about 14 to about 24 carbon atoms in the chainportion of the fatty acid, with about 16 to about 18 carbon atoms beingparticularly preferred in certain embodiments. In certain embodimentsthe fatty acid carbon chain may comprise an odd number of carbon atoms,however, an even number of carbon atoms in the chain may be preferred incertain embodiments. A fatty acid comprising only single bonds in itscarbon chain is called saturated, while a fatty acid comprising at leastone double bond in its chain is called unsaturated.

Specific fatty acids include, but are not limited to, linoleic acid,oleic acid, palmitic acid, linolenic acid, stearic acid, lauric acid,myristic acid, arachidic acid, palmitoleic acid, arachidonic acidricinoleic acid, tuberculosteric acid, lactobacillic acid. An acidicgroup of one or more fatty acids is covalently bonded to one or morehydroxyl groups of a glycerol. Thus, a monoglyceride comprises aglycerol and one fatty acid, a diglyceride comprises a glycerol and twofatty acids, and a triglyceride comprises a glycerol and three fattyacids.

A phospholipid generally comprises either glycerol or an sphingosinemoiety, an ionic phosphate group to produce an amphipathic compound, andone or more fatty acids. Types of phospholipids include, for example,phophoglycerides, wherein a phosphate group is linked to the firstcarbon of glycerol of a diglyceride, and sphingophospholipids (e.g.,sphingomyelin), wherein a phosphate group is esterified to a sphingosineamino alcohol. Another example of a sphingophospholipid is a sulfatide,which comprises an ionic sulfate group that makes the moleculeamphipathic. A phopholipid may, of course, comprise further chemicalgroups, such as for example, an alcohol attached to the phosphate group.Examples of such alcohol groups include serine, ethanolamine, choline,glycerol and inositol. Thus, specific phosphoglycerides include aphosphatidyl serine, a phosphatidyl ethanolamine, a phosphatidylcholine, a phosphatidyl glycerol or a phosphatidyl inositol. Otherphospholipids include a phosphatidic acid or a diacetyl phosphate. Inone aspect, a phosphatidylcholine comprises adioleoylphosphatidylcholine (a.k.a. cardiolipin), an eggphosphatidylcholine, a dipalmitoyl phosphalidycholine, a monomyristoylphosphatidylcholine, a monopalmitoyl phosphatidylcholine, a monostearoylphosphatidylcholine, a monooleoyl phosphatidylcholine, a dibutroylphosphatidylcholine, a divaleroyl phosphatidylcholine, a dicaproylphosphatidylcholine, a diheptanoyl phosphatidylcholine, a dicapryloylphosphatidylcholine or a distearoyl phosphatidylcholine.

A glycolipid is related to a sphinogophospholipid, but comprises acarbohydrate group rather than a phosphate group attached to a primaryhydroxyl group of the sphingosine. A type of glycolipid called acerebroside comprises one sugar group (e.g., a glucose or galactose)attached to the primary hydroxyl group. Another example of a glycolipidis a ganglioside (e.g., a monosialoganglioside, a GM1), which comprisesabout 2, about 3, about 4, about 5, about 6, to about 7 or so sugargroups, that may be in a branched chain, attached to the primaryhydroxyl group. In other embodiments, the glycolipid is a ceramide(e.g., lactosylceramide).

A steroid is a four-membered ring system derivative of a phenanthrene.Steroids often possess regulatory functions in cells, tissues andorganisms, and include, for example, hormones and related compounds inthe progestagen (e.g., progesterone), glucocoricoid (e.g., cortisol),mineralocorticoid (e.g., aldosterone), androgen (e.g., testosterone) andestrogen (e.g., estrone) families. Cholesterol is another example of asteroid, and generally serves structural rather than regulatoryfunctions. Vitamin D is another example of a sterol, and is involved incalcium absorption from the intestine.

A terpene is a lipid comprising one or more five carbon isoprene groups.Terpenes have various biological functions, and include, for example andwithout limitation, vitamin A, coenyzme Q and carotenoids (e.g.,lycopene and β-carotene).

B. Charged and Neutral Lipid Compositions

In certain embodiments, a lipid component of a composition in accordancewith the present disclosure may be uncharged or primarily uncharged. Inone embodiment, a lipid component of a composition comprises one or moreneutral lipids. In another aspect, a lipid component of a compositionmay be substantially free of anionic and cationic lipids, such ascertain phospholipids (e.g., phosphatidyl choline) and cholesterol. Incertain aspects, a lipid component of an uncharged or primarilyuncharged lipid composition comprises about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 80%, about 85%, about 95%, about96%, about 97%, about 98%, about 99% or 100% lipids without a charge,substantially uncharged lipid(s), and/or a lipid mixture with equalnumbers of positive and negative charges.

In other aspects, a lipid composition may be charged. For example,charged phospholipids may be used for preparing a lipid compositionaccording to the present invention and can carry a net positive chargeor a net negative charge. In a non-limiting example, diacetyl phosphatecan be employed to confer a negative charge on the lipid composition,and stearylamine can be used to confer a positive charge on the lipidcomposition.

C. Making Lipids

Lipids can be obtained from natural sources, commercial sources orchemically synthesized, as would be known to one of ordinary skill inthe art. For example, phospholipids can be from natural sources, such asegg or soybean phosphatidylcholine, brain phosphatidic acid, brain orplant phosphatidylinositol, heart cardiolipin and plant or bacterialphosphatidylethanolamine. In another example, lipids suitable for useaccording to the present invention can be obtained from commercialsources. For example, dimyristyl phosphatidylcholine (“DMPC”) can beobtained from Sigma Chemical Co., dicetyl phosphate (“DCP”) may beobtained from K & K Laboratories (Plainview, N.Y.); cholesterol (“Chol”)may be obtained from Calbiochem-Behring; dimyristyl phosphatidylglycerol(“DMPG”) and other lipids known to those of skill in the art may beobtained from Avanti Polar Lipids, Inc. (Birmingham, Ala.). In certainembodiments, stock solutions of lipids in chloroform orchloroform/methanol can be stored at about −20° C. Preferably,chloroform is used as the only solvent since it is more readilyevaporated than methanol, allowing for more expedient lipid recovery.

D. Lipid Composition Structures

In one preferred embodiment of the invention, the drugs may beassociated with one or more lipids, instead of or in addition to, thefatty-acid. In accordance with this aspect of the disclosure, a drugassociated with a lipid may be dispersed in a solution containing alipid, dissolved with a lipid, emulsified with a lipid, mixed with alipid, combined with a lipid, covalently bonded to a lipid, contained asa suspension in a lipid, contained or complexed with a micelle orliposome, or otherwise associated with a lipid or lipid structure. Alipid or lipid/chimeric polypeptide associated composition of thepresent invention is not limited to any particular structure. Forexample, they may also simply be interspersed in a solution, possiblyforming aggregates which are not uniform in either size or shape. Inanother example, they may be present in a bilayer structure, asmicelles, or with a “collapsed” structure. In another non-limitingexample, a lipofectamine (Gibco BRL)-chimeric polypeptide or Superfect(Qiagen)-chimeric polypeptide complex is also contemplated.

In accordance with certain aspects of the present disclosure, a fattyacid- or lipid-containing composition may comprise about 1%, about 2%,about 3%, about 4% about 5%, about 6%, about 7%, about 8%, about 9%,about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%,about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%,about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%,about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about55%, about 56%, about 57%, about 58%, about 59%, about 60%, 61%, about62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%,about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%,about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about88%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%,about 96%, about 97%, about 98%, about 99%, about 100%, or any rangederivable therein, of a particular lipid, lipid type or fatty acid, incombination with one or more therapeutic components such as a drug,biologic agent, or other therapeutic material disclosed herein or aswould be known to one of skill in the art. In a non-limiting example, alipid composition may comprise about 10% to about 20% neutral lipids,and about 13% to about 84% of a tocopherol such as gamma-tocopherol, andabout 1% cholesterol. Thus, it is contemplated that lipid compositionsof the present invention may comprise any of the lipids, lipid types orother components in any combination or percentage range.

The compositions according to the present disclosure may also compriseantibiotics as the drug, or in combination with one or more drugs, e.g.,in combination with tocopherol. Antibiotics suitable for use accordingto the invention are selected from the group including but not limitedto amoxycillin, ampicillin, penicillin, clavulanic acid, aztreonam,imipenem, streptomycin, gentamicin, vancomycin, clindamycin, ephalothin,erythromycin, polymyxin, bacitracin, amphotericin, nystatin, rifampicin,tetracycline, coxycycline, chloramphenicol, and zithromycin.

Compositions according to the invention may also contain a “gellingagent” in combination with the drug or biologically active agent andlipid. The gelling agent may be selected from the group including butnot limited to hydroxyethyl cellulose (HEC), hydroxymethylcellulose(HMC), Natrasol®, pectines, agar, alginic acid and its salts, guar gum,pectin, polyvinyl alcohol, polyethylene oxide, cellulose and itsderivatives, propylene carbonate, polyethylene glycol, hexylene glycolsodium carboxymethylcellulose, polyacrylates,polyoxyethylene-polyoxypropylene block copolymers, pluronics, wood waxalcohols, tyloxapol (a nonionic surfactant oligomer), proteins andsugars.

IV. Therapeutic Treatment

The formulations of water-insoluble and substantially water-insolublecompounds may be used, in combination with the nebulizer systemsdescribed herein, in order to administer a therapeutically effectiveamount of one or more drugs or biological agents as an aerosolizedmixture. Preferably, and in accordance with an aspect of the presentdisclosure, the formulations comprising one or more water-insolublecompounds and a lipid can be administered using a nebulizer as describedherein for the treatment of one or more pulmonary diseases. Pulmonarydiseases and disorders which may be treatable using the compositions,formulations, methods, and apparatus/systems of the present disclosureinclude but are not limited to asthma; alpha-1 antitrypsin deficiency(AAT Deficiency); dust-related pulmonary and lung diseases anddisorders, including asbestosis; avian flu; bronchitis, including acutebronchitis; bronchiectasis; bronchopulmonary dysplasia (BPD); chroniccough; chronic obstructive pulmonary diseases and disorders; the commoncold; chronic obstructive pulmonary disorder (COPD); croup; cysticfibrosis (CF); emphysema; farmer's lung; influenza; hantavirus; rhinitis(hay fever); histoplasmosis; interstitial lung disease; legionellosis(Legionnaire's disease); lung cancer (including both small cell, largecell and mixed small cell/large cell carcinoma); lung damage resultantfrom inhalation of smoke and heat; inflammation and lung damageresultant from inhalation of chemicals; lymphangioleiomyomatosis (LAM);occupational lung disease; pleurisy; pneumonia; pneumothorax; pulmonaryembolus; pulmonary fibrosis; pulmonary hypertension; respiratorydistress syndrome; respiratory syncytial virus (RSV); sarcoidosis;severe acute respiratory syndrome (SARS); sleep apnea; and tuberculosis,as well as two or more such diseases or disorders exhibiting themselvessimultaneously. In accordance with one embodiment of the presentdisclosure, the preferred pulmonary disorder to be treated is lungdamage resultant from inhalation of heat and smoke. In accordance with afurther embodiment of the present disclosure, the pulmonary disorder tobe treated is bronchitis.

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventors to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the scope of theinvention.

EXAMPLES Example 1 Comparative Example

Two ultrasound nebulizers, DeVilbiss Ultra-Neb 99 (available fromSunrise Medical, Respiratory Products Div., Somerset, Pa.) andAeroneb-Pro® (available from Aerogen, Inc., now Nektar Therapeutics,Mountain View, Calif.) and an AirLife™ jet nebulizer (Cardinal Health,Inc., Dublin, Ohio) were selected for testing tocopherol nebulization.The viscose tocopherol preparation (neat tocopherol) would not nebulizewith these devices, regardless of their manipulation or air flowadjustment. Tocopherol was then dissolved into an essential fatty acidrich flax seed oil (linseed oil) preparation (comprising a variety offatty acids, including linolenic acid, linoleic acid, oleic acid,stearic acid, and palmitic acid) at a concentration of 8.3% w/w. Thismuch less viscose mixture was then introduced into the selectednebulizers above and again tested. As before, the tocopherol-fatty acidmixture could not be nebulized using these commercially-availabledevices. It is apparent from these tests that the existing, commerciallyavailable nebulizing apparatus are not designed for, and are largelyincapable of, nebulizing viscose liquids into droplets of an appropriatesize suitable for pulmonary delivery.

Example 2 Measured Air Flow of Nebulizer Nozzles of the PresentDisclosure

Table 1 demonstrates the measured air flow of selected micro-channelnozzle configurations whereby a selected fluid nozzle 14 is surroundedby an air delivery tube 12. Nebulizing air in this example is deliveredat 50 psi. The free air space surrounding the fluid delivery tube 14enclosed by the air delivery tube 12 provides for the calculated “SQinch Neb Free Air Opening”. The actual nebulized air flow rate “Neb AirFlow cc/sec” is related to the free air opening size however, the amountof air flowing through a given free air space is also influenced by theresistance offered by the surfaces of the delivery tubes 14 and 12 incontact with the air and may not be linear in function.

TABLE 1 Approximate Air flow rates with various nozzle combinationconfigurations. Fluid Neb Air SQ inch Neb *Neb Air Gauge Size O.D. I.D.Free Air Flow Config. Fluid/Air “1 O.D.” “2 I.D.” Opening cc/sec A 22/16.0280 .0470 .00111920 175 B 22/18 .0280 .0330 .00023955 30 C 25/16 .0200.0470 .00142079 267 D 25/18 .0200 .0330 .00054114 110 E 25/20 .0200.0230 .00001013 40 F 27/16 .0160 .0470 .00153389 300 G 27/18 .0160 .0330.00065424 140 H 27/20 .0160 .0230 .00021441 60 I 27/22 .0160 .0170.00000259 15 J 30/16 .0120 .0470 .00162185 320 K 30/18 .0120 .0330.00074220 110 L 30/20 .0120 .0230 .00030238 95 M 30/22 .0120 .0160.00008796 45 *Air pressure at 50 PSI

Table 2 demonstrates the fluid delivery rates of various viscosityfluids though various fluid delivery tubes 14 (FIG. 1A-1C) at variousfluid pressures. The actual fluid flow rate is related to the deliverytube internal diameter size however, the amount of fluid flowing througha given fluid opening is also influenced by the resistance offered bythe surfaces of the delivery tube 14 in contact with the fluid as wellas the surface tension of the fluid to be delivered and may not belinear in function. Configurations are based on the nozzleconfigurations described in Table 1, above.

TABLE 2 Approximate Fluid Flow Rates at Various Fluid Pressures WithVarious Fluid Nozzles. Fluid Flow rate Flow Rate Fluid Pres- 100% 0.9%Flow Rate Nozzle Internal sure ETOH Saline Flax Oil Config. G Sizediameter PSI cc/sec cc/sec cc/sec A, B 22 .0160 5 0.125 0.110 0.0034722A, B 22 .0160 10 0.333 0.300 0.0073529 A, B 22 .0160 20 0.435 0.4000.0195744 A, B 22 .0160 30 1.000 0.857 0.0241935 C-E 25 .0100 5 0.02330.0221 C-E 25 .0100 10 0.050 0.0487 C-E 25 .0100 20 0.111 0.1071 C-E 25.0100 30 0.200 0.1866 0.0038265 F-I 27 .0080 5 0.010 0.0092 F-I 27 .008010 0.023 0.0189 F-I 27 .0080 20 0.045 0.0396 F-I 27 .0080 30 0.68 0.06180.0023585 J-M 30 .0060 5 0.00336 0.003195 J-M 30 .0060 10 0.006980.006662 J-M 30 .0060 20 0.01376 0.013102 J-M 30 .0060 30 0.0200270.019032 0.0006527

Table 2 demonstrates the measured air flow of selected micro-channelnozzle configurations whereby a selected fluid nozzle 14 is surroundedby an air delivery tube 12. Nebulizing air in this example is deliveredat about 50 psi. The free air space 16 surrounding the fluid deliverytube 14 enclosed by the air delivery tube 12 provides for the calculated“SQ inch Neb Free Air Opening”. The actual nebulized air flow rate “NebAir Flow cc/sec” is related to the free air opening size however, theamount of air flowing through a given free air space is also influencedby the resistance offered by the surfaces of the delivery tubes 14 and12 in contact with the air and may not be linear in function.

Example 3

In this example, and as illustrated in Table 3, the preferredconfigurations derived from Tables 1 and 2 are demonstrated. This is byway to show the preferred nebulizing air flow rates of the combinationssited but does not limit the arrangements of various combinations offluid and air tube sizes that achieve the preferred embodiment. Thepreferred free nebulizing air open area ranges from about 0.000009 to0.001 square inches. Nebulizing air pressures can be lowered ininstances were there is a larger free air opening in order to bring thenebulizing air flow rate into the preferred range of ≧100 cc/second.Nebulizing air pressures, fluid pressures, micro-channel fluid tube sizeand the viscosity of the fluid to be nebulized are selected from a rangeof components of fluid delivery tubes 1 and air delivery tubes 2 toachieve the preferred air and fluid flow rates and the nebulized dropletsize within the preferred air volume to viscous liquid volume ratio ofless than about 60,000:1.

TABLE 3 Preferred Nozzle Configurations *Neb Air Gauge Size Flow Config.Fluid/Air cc/sec B 22/18 30 D 25/18 110 E 25/20 40 H 27/20 60 K 30/18110 L 30/20 95 M 30/22 45 *Air pressure at 50 PSI

Example 4

Animals:

Adult female sheep were cared for in the Investigative Intensive CareUnit at the University of Texas, Galveston Branch. The experimentalprocedure was approved by the Animal Care and Use Committee of theUniversity of Texas Medical Branch. The National Institutes of Healthand American Physiological Society guidelines for animal care werestrictly followed. The Investigative Intensive Care unit is accreditedby The Association for the Assessment and Accreditation of LaboratoryAnimal Care International.

Animal Model:

Sheep (30-40 kg) were surgically prepared, as described by Enkhbaatar,P. K., et al. [Am. J. Physiol. Regul. Inter. Comp. Physiol., Vol.285(2): R366-R372 (2003)]. A Swan-Ganz thermal dilution catheter (model93A-1317-F, Edwards Critical Care Division, Irvine, Calif.) was insertedthrough the right external jugular vein for the measurement of the corebody temperature to evaluate blood gas and the fluid resuscitations. Anarterial catheter was inserted into the right femoral artery (16 gauge,24 in., Intracath, Becton Dickinson, Sandy, Utah) for the measurement ofarterial blood gas. To evaluate changes in lung lymph flow, an efferentlymph vessel from the caudal mediastinal lymph node was cannulated(Silastic catheter 0.025-in ID, 0.047-in OD; Dow Corning, Midland,Mich.) according to a modification of the technique described by Stauband colleagues [Staub, N., et al., J. Surg. Res., Vol. 19, pp. 315-320(1975); Traber, D., et al., J. Appl. Physiol., Vol. 54, pp. 1167-1171(1983)]. After a 7-day recovery period, the sheep were deeplyanesthetized with halothane and were given a burn (40% total bodysurface area [TBSA], third degree) and inhalation injury (48 breaths ofcotton smoke, <40° C.). After burn/smoke injury, all sheep were placedon a ventilator with positive end-expiratory pressure set to 5 cm H₂Oand tidal volume maintained at 15 mL/kg. The latter tidal volume isequal to about 10 ml/kg in humans due to the large dead space of sheep[Melo, V., et al., Anesthesiology, Vol. 97, pp. 671-681 (2002)]. Allanimals were given fluid resuscitation with Ringer's solution strictlyaccording to the Parkland formula (4 mL/kg % TBSA burned/24 hr). Theexperiment was continued for 48 hr.

Animal Grouping:

The animals were randomized into 3 groups as follows: (1) a Vitamin Enebulization group (B&S, Vitamin E, n=6). To achieve the desiredparticle size, 1 gram mixed tocopherols [1000 mg Decanox™ MTS-90G (94mg/g alpha tocopherol, 15 mg/g beta-tocopherol, 604 mg/ggamma-tocopherol, 201 mg/g delta tocopherol for a total of 914 mg/gtotal mixed tocopherols), purchased from Daniels Midland Co., Decatur,Ill.] was added to 11 grams of linseed oil (flax oil) and mixed for 3hours to make an 8.3% (w/w) solution of tocopherol in linseed oil beforestarting the nebulization, using a nebulizer as described with thepresent disclosure. Nebulization was started 1 hour after the combinedburn and smoke inhalation injury, and repeated every 12 h. (2) A salinenebulization group (B&S, Saline, n=6): injured and nebulized with 15 mLsaline at the same intervals as the vitamin E treatment. (3) A shamgroup (Sham, n=5): cared for similarly to the other groups, that is,given the same amount of fluid as burned animals, placed on aventilator, and studied for 48 h but not injured or treated with salineor tocopherol.

Burn and Smoke Inhalation Injury:

The protocol followed was similar to that described in the art [Kimura,R., et al., J. Appl. Physiol., Vol. 64(3): pp. 1107-1113 (1988);Enkhbaatar, P., et al., Am. J. Physiol. Regal. Integr. Comp. Physiol.,Vol. 285(2): pp. R366R-372 (2003)]. Briefly, under induction ofanesthesia with 10 mg/kg ketamine (Ketalar, Parke-Davis, Morris Plains,N.J.), a tracheotomy was performed, and a cuffed tracheostomy tube(10-mm diameter; Sheiley, Irvine, Calif.) was inserted. Anesthesia wasmaintained with halothane. Using a Bunsen burner, a third-degree flameburn of 20% of the total body surface area was made on 1 flank of thesubject. Thereafter, inhalation injury was induced while the sheep wasin the prone position as described previously [id.]. A modified beesmoker was filled with 50 g burning cotton toweling and was connected tothe tracheostomy tube via a modified endothoracheal tube containing anindwelling thermistor from a Swan-Ganz catheter. During the insufflationprocedures, the temperature of the smoke did not exceed 40° C. The sheepwere insufflated with a total of 48 breaths of cotton smoke. After smokeinsufflation, another 20% total body surface area, third-degree burn,was made on the contralateral flank.

Resuscitation Protocol:

The protocol for subject resuscitation has been described previously inthe art [id.]. Briefly, immediately following the injury, anesthesia wasdiscontinued and the animals were allowed to awaken and weremechanically ventilated with a Servo ventilator (model 900C,Simens-Elena, Solna, Sweden) throughout the next 48 h experimentalperiod. Ventilation was performed with a positive end-expiratorypressure (PEEP) of 5 cm H₂O and a tidal volume of 15 mL/kg. Therespiratory rate was set to maintain normocapnia. For the first 3 hpost-injury, all animals received an inspired oxygen concentration(FiO₂) of 100% to expedite the removal of carbon monoxide (CO);thereafter, the FiO₂ was adjusted to maintain the arterial oxygensaturation to be greater than 90%. These respiratory settings allowedrapid carboxyhemoglobin clearance after smoke inhalation.

During the experiment, fluid resuscitation was performed with Ringerlactate solution following the formula (4 mL/% burn surface area/kg bodyweight for the first 24 h and 2 mL/% burned surface area/kg body weightper day for the next 48 hours). During this experimental period, theanimals were allowed free access to food, but not to water, to allowaccurate determination of fluid balance.

Concurrent cutaneous burn and smoke inhalation (B&S) injuries have grossevidence of lung oxidant injury. Additionally, inflammatory blood cells,neutrophil granulocytes, infiltrate the lung tissues causing edema,swelling and additional tissue damage from products of the inflammatorycells. Neutrophil infiltrate is an integral part of the formation orobstructive airway cast formation following inhalation injury.Obstructive airway casts is the major cause of pulmonary obstructionfollowing inhalation injury. Pulmonary obstruction reduces the flow ofair in and out of the lung and is the primary cause of death followinginhalation injury. Therefore, we hypothesized that direct lunginsufflation by nebulization of gamma-tocopherol (γ-T), a potentreactive oxygen and nitrogen scavenger, would attenuate this injury byreducing oxidative stress. Additionally, flax seed oil as used in thepresent examples is made up of about 69% short chain essential fattyacids, which also have been shown to exhibit anti-inflammatoryproperties, and was used as a carrier for an 8.3% solution of mixedtocopherols.

Acute lung injury was induced in adult female sheep by smoke inhalation(48 breaths of cotton smoke, <40° C.) with concurrent 40% TBSA 3rddegree cutaneous burn under deep anesthesia. Sheep (35.4±1.0 kg) weredivided into 4 groups: 1) sham (not injured, nebulized flax oil (FO))n=4; 2) saline (injured, nebulized saline, n=6); 3) FO (injured,nebulized FO, n=4); and 4) gamma-tocopherol+FO (injured, nebulizedFO-gT, n=4). Insufflation by nebulization was started 1 h post-injuryand 22 ml FO with or without g-T (mixed natural tocopherols, 528 mg/48h) was continuously delivered into the pulmonary system for 48 hoursusing the subject lipid nebulization system using the control system asshown in FIG. 4 with a nebulizing nozzle in accordance with FIG. 1A-1C,and configured as described in Table 1, Configuration H, of Example 1.

Cardiopulmonary variables were unchanged in sham animals receiving flaxseed oil over 48 hours and flax seed oil with 8.3% mixed tocopherols(FIG. 7A). Combined burn and smoke inhalation injury caused severepulmonary dysfunction evidenced by deteriorated pulmonary gas exchange(FIG. 7A), increased pulmonary vascular permeability (FIGS. 7B and 7C),and increased ventilatory pressures (FIG. 7D) were observed. Thesechanges were associated with decreased lung gT levels. FO nebulizationsignificantly improved the pathological changes seen in the saline group(FIGS. 7A-D). Gamma-tocopherol addition further improved pulmonaryfunction (FIGS. 6A-D) along with 100-fold elevation in the lung γ-Tconcentrations (18.0±7.6 vs. 0.017±0.005 in saline group).

Pulmonary administration by nebulization of flax seed oil did notinterfere with gas exchange or other pulmonary function whenadministered at a rate of ≦0.5 cc per hour in uninjured sham animals.All (6/6) saline nebulized control animals (FIG. 7A) deteriorated intorespiratory distress (ARDS defined as a blood PO₂ ratio to inspired O₂concentration of ≦200; PO₂:FiO₂ or P/F Ratio≦200) within 24 hours of thecombined burn and smoke inhalation injury. None (0/4) of the animalsreceiving nebulized flax oil with 8.3% mixed tocopherols administered at0.3 to 0.5 cc/hour deteriorated into ARDS. All physiological parameters(FIGS. 7A-D) were significantly improved in the insufflated flaxoil/tocopherol group as compared with the saline nebulization group.

Histological examination of lung and burn wound tissues was performedduring animal sacrifice necropsy at 48 hours post injury. Profoundinflammatory cell infiltrate of neutrophils was noted in the B&S Salinecontrol group in both the lung and burn wound tissues. Neutrophilinfiltrate was noted in airway casts following smoke inhalation injuryin the B&S Saline control group. A marked reduction in inflammatory cellneutrophil infiltrate was noted in the flax (FO) group and thegamma-tocopherol+flax (FO-gT) group. These observations along with thephysiological measurements demonstrate that inspired flax (FO) and thegamma-tocopherol+flax (FO-gT) reduces the local and systemicinflammatory response following a smoke inhalation injury.

Example 5 Comparison of Lipid Carrier Compositions

Two medicinal lipid compositions were prepared, and compared using thetechniques described in the examples above. The first compositioncomprised tocopherol and flax seed oil as a carrier, wherein the flaxseed oil comprised at least five (5) fatty acids, as shown in Table 4,below. The second, comparative composition comprised tocopherol and a“special” purified fatty-acid mixture (available from Nu-Chek Prep,Inc., Elysin, Minn.) having a high (greater than about 50 wt. %)linolenic acid content. The comparative compositions are shown in Table4. Initial tests seem to suggest that the use of the “special” mixturewith the high omega-3-linolenic acid content may be even moreadvantageously beneficial in reducing inflammatory response in the lungsof a patient, following lung injury.

TABLE 4 Comparison of standard Flax Seed oil and Purified “Special” Oilfrom Nu-Chek Prep., Inc. Linolenic Linoleic Stearic Palmitic Acid AcidOleic Acid Acid Acid Oil (C-18) (C-18) (C-18) (C-18) (C-16) Flax SeedOil 47 wt. % 24 wt. % 19 wt. % 3 wt. % 6 wt. % (Linseed) Purified 68 wt.% 29 wt. %  3 wt. % — — “Special” Oil

Example 6 Comparison of Lipid Carrier Compositions

In this example, nebulized fatty acid droplets (nebulized in accordancewith aspects and systems of the present disclosure) were impacted on acounting slide and examined, as illustrated in FIG. 8. The picturedflattened, semi-hemispherical droplets in FIG. 8 are more than twice thediameter of the spherical droplets before impact. Oils containing largeamounts of linolenic and linoleic fatty acids tend to “flow out” asthese fatty acids are natural wetting agents. The droplet population iswithin the target size distribution of 1-5 μm Mass Median AerodynamicDiameter (MAMD). The small in-flight droplet size creates imagingdifficulties with regard to depth of field at required magnificationsfor visualization. It is worth noting that the difficulty in measurementof droplets≦5 μm has been acknowledged by many others in the art.

Example 7 Additional Aerosol Formulation Delivery Studies

Animals/Animal Model

Twenty-four adult female sheep were cared for, prepared, and maintainedas described in Example 4, above.

Experimental Design

The sheep were randomly assigned to one of the following four groups: 1)Sham and nebulized with flax oil (FO) (not injured, FO-nebulized, n=6);2) Saline (injured, saline-nebulized, n=6); 3) FO (injured,FO-nebulized, n=6); 4) γ-T+FO (injured, FO+γ-T-nebulized, n=6). Shamanimals received no injury but were surgically prepared like treatedanimals, placed on a ventilator, and given fluid resuscitation andnebulized with 24 ml FO over 47 hr. Saline animals were nebulized with24 ml of 0.9% NaCl over 47 hr after injury. FO animals were nebulized 24ml FO over 47 hr after injury. γ-T+FO animals were nebulized 24 mlγ-T+FO mixture solution (γ-T:1220 mg) over 47 hr after injury.

Aerosol Delivery & Material.

The novel, viscous lipid formulation nebulization nozzle and controlsystem described herein was adapted to a Siemans® 900c servo ventilator(Siemans-Elema AB, Sweden), as shown and discussed generally in relationto FIGS. 1-6B, above. Briefly, for purposes of this particularexperiment, the nebulizing nozzle (FIG. 1B) was fabricated fromhypodermic needle stock material with a center fluid delivery tube andan outer air delivery tube, in accordance with aspects of the presentdisclosure. For the purpose of calibration, calibrated blood countingslides were waved through the ventilator inspiratory airflow containingnebulized flax oil formulations allowing droplets to impact onto theslide. The slides were then observed under light microscopy (200×) andsized visually and counted. The vast majority (100:1) of observedimpacted deformed and flattened droplets larger than 2 μm and smallerthan 10 μm in diameter were conservatively considered to be in the 2-5μm spherical range.

The output end of the nebulizing nozzle is positioned in the center offlow within the “Y” connector of the ventilator circuit immediatelyadjacent to and directed toward the tracheostomy tube connector. Thenozzle is fed from an air/liquid flow control system adapted to andcontrolled by the electronic output of the 900c ventilator. Theoxygen-air mixer blending the inspired air for the ventilator is tappedto provide the nebulizing air supply providing the nebulization air atthe same FiO₂ as the ventilator air FiO₂. The control cycle timers areprogrammed to provide 1.0 second of air flow and 0.4 seconds of fluidflow with each inspiration cycle. The fluid flow and subsequentnebulization is configured to occur in the first 0.6 seconds ofinspiration thereby providing the nebulized droplets at the beginning ofthe inspired air flow into the lungs. This nebulizer configurationprovides an additional inspired air flow volume of 50 mL/second/breathwhich was deducted from the total tidal volume.

Cold pressed, filtered flax seed oil (Spectrum Organic Products LLC,Melville, N.Y.) was used alone and as a carrier for 8.3% solution w/w ofmixed tocopherols (Decanox™ MTS-90G, a gift from Dr. Brent Flickinger,Archer Daniels Midland Co., Decatur, Ill.) which was measured to containγ-T (610 mg/g) and, α-T (91 mg/g). The flax oil mixtures were sterilefiltered through a 0.22 μm pore filter prior to use. Flax oil alone orflax oil containing 8.3% Decanox™ was administered by continuous pulsenebulization synchronized with the inspiration cycle at a rate of about0.45-0.5 ml/hour or 11-12 ml/24 hours. Direct lung tissue γ-Tconcentration measurements (FIG. 9) demonstrated that the aerosolizedmaterial was deposited into the lung.

Measured Variables

Arterial and mixed venous blood samples were taken at different timepoints for measurement of blood gases (IL GEM Premier 3000 Blood GasAnalyzer; GMI, Minnesota). PaO₂/F_(i)O₂ ratio was measured to helpassess pulmonary gas exchange. The pulmonary microvascular fluid fluxwas evaluated by measuring the lung lymph flow. Sheep were sacrificedunder deep halothane anesthesia 48 hr after injury. The right lung wasthen removed, and a 1-cm-thick section was taken from the middle of thelower lobe, injected with 10% formalin, and immersed in formalin. Fourtissue samples were taken at predetermined sites for histologicalexamination. Fixed samples were embedded in paraffin, sectioned at 4 μm,and stained with hematoxylin and eosin. A pathologist without knowledgeof the group assignments evaluated the lung histology. Levels of airwayobstruction were obtained with a standardized protocol. Fifteen bronchiwere investigated, and the percentage of area obstructed by the cast wasestimated (0%-100%). The remaining lower one-half of the right lowerlobe was used for the determination of bloodless wet-to-dry weightratio. Pulmonary shunt fraction (Qs/Qt) was calculated using standardequations.

γ-tocopherol Measurement

A modification of the method by Podda and colleagues [Podda, M., et al.,J. Lipid Res., Vol. 37, pp. 893-901 (1996)] was used for α- and γ-Tanalyses, as described previously [Morita, N., et al., SHOCK, Vol.25(3), pp. 277-282 (2006)]. Briefly, tissue (˜50 mg) or plasma (100 μL)was saponified with alcoholic KOH, extracted with hexane, the extractdried under nitrogen, the residue resuspended in 1:1 ethanol-methanol,then injected into an HPLC system. Tocopherols were detected using anelectrochemical detector, and quantitated by comparison to authenticstandards. Tissue γ-T levels were adjusted by the wet and dry ratio tocorrect for the weight change caused by the inflammation and edema.

Malondialdehyde Measurement

Malondialdehyde (MDA) concentrations were utilized to estimate the lipidperoxidation in the lung and were measured as thiobarbituric acidreactive material. Lung tissue MDA levels were quantified with acommercially available assay (Northwest Life Science Specialties,Vancouver, Wash.). The level of lipid peroxides is expressed as MDA permilligram protein, measured using a commercially available assay (FlukaBioChemika, Buchs, Switzerland).

3-Nitotyrosine Measurement

3-Nitrotyrosine concentration, an index of the nitrosylation ofproteins, was determined by enzyme-linked immunosorbent assay (ELISA).After the study, samples of lung tissue were collected and homogenatesprepared after adding 2 mL of the 10× diluted halogenations buffer(1:10; Cayman Chemical, Ann Arbor, Mich.) containing 250 mM Tris-HCL (pH7.4), 10 mM EDTA, and 10 mMethyleneglycol-bis(-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA).The supernatant was obtained by centrifugation (10,000×g at 4° C.) for15 min and the supernatant used for assessment. We used the Hycutbiotechnology 3-NT solid phase ELISA (Cell Sciences Inc., Canton, Mass.)according to the manufacturer's protocol.

Immunohistochemistry of PARP Activity

For the immunohistochemical detection of poly(ADP-ribose), mousemonoclonal anti-PAR antibody (Calbiochem, San Diego, Calif., USA)(1:1000, overnight, 4° C.) was used after antigen retrieval. Secondarylabeling was achieved by using biotinylated horse anti-mouse antibody(Vector Laboratories, Burlingame, Calif., USA) (30 min roomtemperature). Horseradish peroxidase-conjugated avidin (30 min, roomtemperature) and brown colored diaminobenzidine (6 min, roomtemperature) was used to visualize the labeling (Vector Laboratories,Burlingame, Calif., USA). The sections were counterstained withhematoxilin (blue color).

The intensity of PAR staining of individual sections was determined by ablinded experimenter according to a semiquantitative PAR-positivityscore from 1-10. (1: no staining, 2: light cytoplasmic staining, 3: fewpositive nuclei, 4: light nuclear staining in approximately 10% ofcells, 5: light nuclear staining in approximately 25% of cells, 6: lightnuclear staining in approximately 50% of cells, 7: strong nuclearstaining in approximately 50% of cells, 8: approximately 75% of thenuclei are positive, 9: approximately 90% of the nuclei are positive,10: few negative cells).

IL-8 and IL-6 mRNA Measurement

Lung tissue was excised at the time of sacrifice and immersed in liquidN₂. Total RNA is obtained using a commercially available total RNApurification kit, Purescript™ (Gentra Systems, Inc., Minneapolis,Minn.). Briefly 100 mg of the freshly frozen lung was lysed andhomogenized using a mortar and pestle with 3 ml of lysis buffercontaining EDTA, citric acid, and SDS according to the manufacturer'sprotocol, except that the homogenized tissue was incubated overnight atroom temperature in the lysis buffer. Precipitation buffer was added andincubated 10 min on ice to precipitate protein and DNA and centrifugedat 3,000 g. The supernatant was placed in 3 ml of isopropanol andcentrifuged at 3,000 g for 5 min. The pellet was washed with 3 ml of 70%ethanol, centrifuged again and air-dried for 10 min. The pellet wasresuspended in DEPC-treated water. Total RNA was quantitatedspectrophotometrically at 260 nm. Quality of the isolated RNA wascontrolled by measuring the ratio of 28s/18s rRNA. Messenger RNA wasisolated from the total RNA by the Straight A's™ mRNA Isolation System(Novagen, Madison, Wis.) purification procedure in which mRNA was firsthybridized to oligo dT coupled magnetic beads, washed, and then elutedto obtain polyadenylated mRNA according to the manufacturer's protocol.First strand cDNA was synthesized by reverse transcription of the mRNAsamples using MMLV-derived reverse transcriptase (Perkin Elmer,Branchburg, N.J.) and random hexamers for priming according to standardtechniques [Sambrook, J., Fritsch, E., and Maniatis, T., “MolecularCloning: A Laboratory Manual.”, Cold Spring Harbor Laboratory Press, NewYork: (1989)]. The cDNA was then used as a template for real-time PCR.Primers and probes were designed using a commercial online primer designprogram (Biosearch Technologies, Inc., Novato, Calif.) (Table 5) andpurchased from the same company. QPCR was performed with a Rotor-Gene™3000 (Corbett Research, San Francisco, Calif.).

TABLE 5 GAPDH, IL-8 and IL-6 Primers and Probes. Probe Conc.Primer Sequence Ov GAPDH  0.5 μM 5′ CGCTCCCATGTTTGTG 3′ Forward Ov GAPDH 0.5 μM 5′ GAGGCATTGCTGACA A 3′ Reverse Ov GAPDH 62.5 μM 5′dTGGGCGTGAACCACGAGAAGTATA 3′ Probe Ov IL-8  0.5 μM 5′GCAACCCTAGACTGCT 3′ Forward Ov IL-8  0.5 μM 5′ CCAGTGAAGAATAAAGAAATCG 3′Reverse Ov IL-8 62.5 μM 5′ TCACGAGTTCCTGTTAACTGTGC 3′ Probe Ov IL-6 0.5 μM 5′ TTGAGGGAAATCAGGAAA 3′ Forward Ov IL-6  0.5 μM 5′GCTGGAGTGGTTATTAGAC 3′ Reverse OV IL-6 62.5 μM 5′TCATGGAGTTGCAGAGCAGTATCA 3′ Probe

The reaction mixtures consisted of dilutions of cDNA from 256 ng of RNA,primer and probe concentrations as indicated (Table 5), and subjected toamplification using a final, optimized concentration of MgCl₂, 0.375 Uof Taq polymerase (AmpliTaq®, Perkin-Elmer) and 0.2 mM dTP's in areaction volume of 15 μl. The mixtures were amplified for 40 cycles at amelting temperature of 95° C. for 10 min, an annealing temperature of55° C. for 10 s, and extension at 60° C. for 45 s. The thresholdamplifications (Ct) for each dilution, and reaction efficiencies weredetermined for each analyte using Rotor-Gene™ software (CorbettResearch). The copy numbers were normalized between samples using GAPDHcopy numbers obtained by determination of GAPDH copy number using anexternal standard constructed from the v-erb gene. All results wereexpressed as copy numbers per μg of total RNA.

Statistical Analysis

Summary statistics of data are expressed as means±standard error of themean. Statistical significance was determined using a two-factoranalysis of variance with repeated measures. The two factors weretreatment and time. Fisher's least significant difference procedure withBonferoni's adjustment for number of comparisons is used for themultiple comparisons (or post-hoc analysis). Effects and interactionswere assessed at the p<0.05 level of significance.

Results

All animals survived the 48 h experimental period after the combinedinjury with 40% TBSA burn and smoke inhalation. There were nostatistically significant differences in the mean arterialcarboxyhemoglobin levels measured immediately after smoke exposurebetween the saline, FO and γ-T+FO groups (67.4%±6%, 77.0%±5% and 68%±7%,respectively). Since vitamin E may decrease platelet adhesion, theclotting time was evaluated. The γ-T treatment did not result in ableeding tendency in any of the groups. The activated clotting time was144±14 s at baseline, 163±3 s at 24 hr, and 160±10 s at 48 hr in theγ-T+FO group and 158±3 s at baseline, 177±10 s at 24 hr, and 183±12 s at48 hr in the nebulized saline group.

The lung contains primarily α-T and relatively low γ-T concentrations insheep. Lung γ-T concentrations were low, and as shown previously, burnand smoke inhalation injury further reduced both α- and γ-Tconcentrations in lung tissue. However, the nebulization significantlyincreased γ-T concentrations in lungs of sheep in the γ-T+FO group. Noincreases were found in plasma γ-T (data not shown) documenting that theγ-T administration is confined to the lung.

Table 6 shows a comparison of effects of FO or γ-T+FO nebulization onpulmonary gas exchange (PaO₂/FiO₂ ratio and Qs/Qt) and pulmonarytransvascular fluid flux (lung lymph flow). There was a significantdecrease in PaO₂/FiO₂ and an increase in pulmonary shunt fraction andlung lymph flow in the saline group resulting from the combined burn andsmoke inhalation injury as compared with the sham group at 24, 36, and48 hr. FO nebulization had a tendency to attenuate the changes seen inanimals with the saline group. As compared with the FO group, the use ofnebulized γ-T+FO significantly improved the PaO₂/FiO₂ at 36 hr and inlung lymph flow at 24 hr and 48 hr. PaO₂/FiO₂ ratio (FIG. 10A) wasmarkedly decreased in animals that were nebulized with saline (injured)as compared with sham animals (uninjured). Nebulization of γ-T+FOattenuated the decrease in this variable. Statistically significantdifferences were observed at 24, 30, 36, 42 and 48 hr compared with thesaline group and at 30, 36, and 42 hr compared with FO group. Anincrease in pulmonary shunt fraction (FIG. 10B) seen in the saline groupwas significantly attenuated by FO nebulization at 48 hr and γ-T+FOnebulization at 36, 42 and 48 hr after the combined injury.

TABLE 6 Pulmonary Gas Exchange and Transvascular Fluid Flux Results.Time (hours, h) 0 6 12 24 36 48 PaO₂/FiO₂ Sham 487 ± 14  575 ± 24  560 ±19 575 ± 14  558 ± 15  560 ± 11  Saline 492 ± 16  467 ± 63  443 ± 35 174± 31* 117 ± 30*  81 ± 17* FO 486 ± 16  525 ± 28  416 ± 59 290 ± 65* 194± 40* 137 ± 24* γ-T + FO 490 ± 13  523 ± 41  440 ± 64  415 ± 55*^(†) 349 ± 48*^(†‡)  270 ± 48*^(†) Qs/Qt Sham 0.19 ± 0.02 0.12 ± 0.01  0.15± 0.01 0.14 ± 0.01 0.13 ± 0.01 0.14 ± 0.01 Saline 0.19 ± 0.04 0.14 ±0.03  0.15 ± 0.02  0.33 ± 0.05*  0.41 ± 0.07* 0.49 ± 0.07 FO 0.17 ± 0.090.14 ± 0.02  0.18 ± 0.03 0.24 ± 0.04  0.30 ± 0.06*   0.33 ± 0.06*^(†)γ-T + FO 0.18 ± 0.01 0.14 ± 0.01  0.20 ± 0.03 0.23 ± 0.05  0.23 ±0.03^(†)  0.26 ± 0.03^(†) Lymph Sham 2.8 ± 0.7 3.5 ± 0.6  3.1 ± 0.6 3.7± 1.4 2.8 ± 0.8 3.2 ± 0.7 Saline 4.8 ± 0.9 11.8 ± 1.8  19.6 ± 5.1 42.7 ±6.0* 46.5 ± 4.1* 48.2 ± 1.9* FO 6.8 ± 2.0 9.4 ± 2.2 14.4 ± 3.4  23.5 ±7.5*^(†)  23.5 ± 7.1*^(†)  26.9 ± 5.9*^(†) γ-T + FO  4.1 ± 1.17 9.9 ±4.3 9.24 ± 3.3  8.6 ± 2.8^(†‡)  9.1 ± 2.5^(†)  10.5 ± 2.2^(†‡) Data areexpressed as means ± SEM. *P < 0.05 vs. Sham. ^(†)P < 0.05 vs. Saline;^(‡)P < 0.05 vs. FO.

Lung lymph flow, a characteristic of pulmonary transvascular fluid flux,was markedly increased in injured, saline nebulized animals comparedwith the sham group (FIG. 11). The lymph flow began to increase 12 hrafter the insult and a peak was observed at 42 hr. However, nebulizationof γ-T+FO reversed this increase in pulmonary transvascular fluid fluxand significant differences were observed between γ-T+FO and Salinegroups at 18, 24, 30, 36, 42 and 48 hr, and γ-T+FO and FO groups at 24and 48 hr after the combined injury. Lung bloodless wet-to-dry weightratio, a measure of lung water content, was significantly increased at48 hr after insult in the saline group as compared with the sham group(FIG. 12A). However, the nebulization of γ-T+FO significantly reducedthis increase.

The airway obstruction score revealed a significant increase in meanobstruction of bronchi (FIG. 12B) in the saline group as compared withthe sham group. Treatment with γ-T+FO nebulization significantly reducedthe obstruction score.

FIG. 13A illustrates the effect of γ-T+FO nebulization onmalondialdehyde concentration which is an index of lipid peroxidation(ROS) in lung tissue. Malondialdehyde concentration was significantlyincreased in the saline group as compared with the sham group.Malondialdehyde levels did not markedly increase in animals treated withγ-T+FO nebulization.

3-Nitrotyrosine is a marker of nitrosative stress, resulting fromreactive nitrogen species (RNS) such as peroxynitrite. Burn and smokeinjury caused a marked increase in lung 3-nitrotyrosine 48 h after theinsults. γ-T+FO nebulization significantly prevented the increase in3-nitrotyrosine (FIG. 13B).

After burn and smoke injury, there was a marked increase in poly(ADP-ribose) reactivity in the Saline and FO groups (FIGS. 14A-14B).Treatment with γ-T+FO nebulization prevented this increase in activity(FIG. 14A). FIG. 14B shows the PAR-positivity score graph whichquantified the degree of poly (ADP-ribose) histochemical stain. Burn andsmoke injury caused a significant increase in lung poly (ADP-ribose)polymerase activity. However, γ-T+FO nebulization significantlyprevented the increase in lung poly (ADP-ribose) polymerase activity.

To determine the pro-inflammatory chemokines, IL-8 and IL-6 mRNA weremeasured in lung tissue (FIG. 15). Burn and smoke injury caused a markedincrease in lung IL-8 and IL-6 mRNA 48 hr after the insults. γ-T+FOnebulization prevented the increase in IL-8 and IL-6 mRNA (FIG. 15).

The invention has been described in the context of preferred and otherembodiments and not every embodiment of the invention has beendescribed. Obvious modifications and alterations to the describedembodiments are available to those of ordinary skill in the art. Thedisclosed and undisclosed embodiments are not intended to limit orrestrict the scope or applicability of the invention conceived of by theApplicants, but rather, in conformity with the patent laws, Applicantsintends to protect all such modifications and improvements to the fullextent that such falls within the scope or range of equivalent of thefollowing claims.

All of the methods, processes and/or apparatus disclosed and claimedherein can be made and executed without undue experimentation in lightof the present disclosure. While the methods, apparatus and processes ofthis invention have been described in terms of preferred embodiments, itwill be apparent to those of skill in the art that variations may beapplied to the methods, apparatus and/or processes and in the steps orin the sequence of steps of the methods described herein withoutdeparting from the concept and scope of the invention. For example,while objects of the present invention have been described as being inspecific spatial relationships such as “parallel to” and “horizontalto”, it is envisioned that such objects can also be at a variety ofangles (e.g., acute, obtuse, or oblique angles) with respect to oneanother without departing from the scope of the present invention. Morespecifically, it will be apparent that certain features which are bothmechanically and functionally related can be substituted for thefeatures described herein while the same or similar results would beachieved. All such similar substitutes and modifications apparent tothose skilled in the art are deemed to be within the scope and conceptof the invention.

Further, any documents to which reference is made in the application forthis patent as well as all references listed in any list of referencesfiled with the application are hereby incorporated by reference.However, to the extent statements might be considered inconsistent withthe patenting of this invention such statements are expressly not to beconsidered as made by the applicant(s).

What is claimed is:
 1. A pulmonary drug delivery system capable ofnebulizing a drug composition comprising a water-insoluble orsubstantially water-insoluble drug and a fatty acid or lipid, the systemcomprising: a carrier gas reservoir; a drug reservoir comprising aflexible bladder that is disposed within the carrier gas reservoir andthat is able to be pressurized by gas within the carrier gas reservoir;a face mask; and a nebulizing nozzle in fluid communication with facemask, the drug reservoir and the carrier gas reservoir, wherein thenebulizing nozzle comprises an outer carrier gas delivery tube spacedapart from and disposed around at least one inner drug delivery tube andwherein the outer carrier gas delivery tube and the at least one innerdrug delivery tube(s) share a longitudinal axis and are togetherdimensioned and disposed such that expulsion of a carrier gas throughthe outer carrier gas delivery tube forms an outer surrounding stream ofcarrier gas that carries an inner flow of fluid from the drug reservoirin a center of carrier gas flow exiting the nebulizing nozzle andthereby produces aerosol drug containing droplets having a particle sizeranging from about 2 μm to about 12 μm in median mass aerodynamic sizethat are delivered to a patient via a mouth and/or nose of the patient.2. The pulmonary drug delivery system of claim 1, further comprising amechanical ventilator.
 3. The pulmonary drug delivery system of claim 2,wherein the mechanical ventilator is configured to synchronize a cyclicnebulization of the aerosol drug containing droplets.
 4. The pulmonarydrug delivery system of claim 1, wherein the nebulizing nozzle canproduce aerosol drug containing droplets having a particle size rangingfrom about 2 μm to about 5 μm.
 5. The pulmonary drug delivery system ofclaim 1, wherein the nozzle has an air volume to nebulized dropletvolume ratio less than about 60,000:1.
 6. The pulmonary drug deliverysystem of claim 5, wherein the nozzle has an air volume to nebulizeddroplet volume ratio less than about 15,000:1.
 7. The pulmonary drugdelivery system of claim 1, wherein an intermediate air space betweenthe outer carrier gas delivery tube and the inner drug delivery tuberanges from about 0.000009 in² to about 0.001 in².
 8. The pulmonary drugdelivery system of claim 7, wherein an intermediate air space betweenthe outer carrier gas delivery tube and the inner drug delivery tuberanges from about 0.00000259 in² to about 0.001 in².
 9. The pulmonarydrug delivery system of claim 1, wherein the outer carrier gas deliverytube has an inner diameter ranging from about 0.01 inches to about 0.05inches.
 10. The pulmonary drug delivery system of claim 1, wherein theouter gas delivery tube is disposed around a plurality of individualinner drug delivery tubes disposed within, and sharing a longitudinalaxis with, the outer gas delivery tube.
 11. The pulmonary drug deliverysystem of claim 10, wherein the plurality of individual drug deliverytubes ranges from 2 to 12 individual drug delivery tubes disposed withinthe outer gas delivery tube.
 12. The pulmonary drug delivery system ofclaim 10, where the relative dimensions of the outer gas delivery tubeand the plurality of individual inner drug delivery tubes produce theaerosol drug containing droplets having a particle size ranging fromabout 2 μm to about 12 μm in median mass aerodynamic size.
 13. Apulmonary drug delivery system capable of nebulizing a drug compositioncomprising a water-insoluble or substantially water-insoluble drug and afatty acid or lipid, the system comprising: a drug reservoir; a carriergas reservoir; a face mask; and a nebulizing nozzle in fluidcommunication with face mask, the drug reservoir and the carrier gasreservoir, wherein the nebulizing nozzle comprises an outer carrier gasdelivery tube spaced apart from and disposed around at least one innerdrug delivery tube and wherein the outer carrier gas delivery tube andthe at least one inner drug delivery tube(s) share a longitudinal axisand are together dimensioned and disposed such that expulsion of acarrier gas through the outer carrier gas delivery tube forms an outersurrounding stream of carrier gas that carries an inner flow of fluidfrom the drug reservoir in a center of carrier gas flow exiting thenebulizing nozzle and thereby produces aerosol drug containing dropletshaving a particle size ranging from about 2 μm to about 12 μm in medianmass aerodynamic size that are delivered to a patient via a mouth and/ornose of the patient, wherein the drug reservoir is disposed within apressurization chamber that is adapted to force delivery of a drug fromthe drug reservoir into the drug delivery tube and wherein thepressurization chamber can be pressurized by the carrier gas reservoir.14. The pulmonary drug delivery system of claim 13, further comprising aspring loaded flow valve positioned and adapted to controlpressurization by the carrier gas reservoir.
 15. The pulmonary drugdelivery system of claim 13, further comprising a spring loaded blow-offvalve positioned and adapted to prevent excess pressure in thepressurization chamber.
 16. A pulmonary drug delivery system capable ofnebulizing a drug composition comprising a water-insoluble orsubstantially water-insoluble drug and a fatty acid or lipid, the systemcomprising: a nebulizing nozzle in fluid communication with a drugreservoir that is disposed within a carrier gas reservoir, wherein thedrug reservoir is adapted to be pressurized by gas within the carriergas reservoir and thereby force delivery of a drug from the drugreservoir into the nebulizing nozzle, and wherein the nebulizing nozzlecomprises an outer carrier gas delivery tube spaced apart from anddisposed around at least one inner drug delivery tube and wherein theouter carrier gas delivery tube and the at least one inner drug deliverytube(s) share a longitudinal axis and are together dimensioned anddisposed such that expulsion of a carrier gas through the outer carriergas delivery tube forms an outer surrounding stream of carrier gas thatcarries an inner flow of fluid from the drug reservoir in a center ofcarrier gas flow exiting the nebulizing nozzle and thereby producesaerosol drug containing droplets having a particle size ranging fromabout 2 μm to about 12 μm in median mass aerodynamic size.
 17. Thepulmonary drug delivery system of claim 16, wherein the drug reservoiris a flexible bladder that is disposed within the carrier gas reservoir.18. The pulmonary drug delivery system of claim 16, further comprising aspring loaded flow valve positioned and adapted to controlpressurization by the carrier gas reservoir.
 19. The pulmonary drugdelivery system of claim 16, further comprising a spring loaded blow-offvalve positioned and adapted to prevent excess pressure in thepressurization chamber.